2012 cell biolabs catalog web
DESCRIPTION
For Sales,QUALITY LAB TECHwww.qlabtechnologies.comTRANSCRIPT
Cell-Based Assays
Ordering Information & Support 4
5
Stem Cell Research 31
Viral Expression 39
microRNA Analysis 73
TRANSFORMATION ADHESION MIGRATION INVASION WOUND HEALING
2 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
TABLE OF CONTENTS
CELL HEALTH CELL HYPOXIA PHAGOCYTOSIS CONTRACTION ANGIOGENESIS
IPS CELL REPROGRAMMING RETROVIRAL EXPRESSION SYSTEMS FEEDER CELLS COLONY FORMATION ASSAYS ALKALINE PHOSPHATASE ASSAYS
ADENO-ASSOCIATED VIRUS (AAV)
ADENOVIRUS LENTIVIRUS RETROVIRUS
PREMADE VIRUSES EXPRESSION SYSTEMS PURIFICATION KITS TITER KITS TRANSDUCTION KITS
PRECURSOR CLONE COLLECTION MAMMALIAN EXPRESSION VECTORS VIRAL EXPRESSION PLASMIDS FUNCTIONAL REPORTER SYSTEM KNOCKDOWN ENHANCER
3
Oxidative Stress / Damage 83
Cell Signaling & Protein Biology 111
Product Index
Worldwide Contacts
151
157
TABLE OF CONTENTS
www.cellbiolabs.com [email protected]
Metabolism Research 129
Pathogen and Toxin Assays 147
PROTEIN OXIDATION/NITRATION LIPID PEROXIDATION DNA/RNA DAMAGE AND REPAIR HYPOXIA ASSAYS REACTIVE OXYGEN SPECIES (ROS) ASSAYS ANTIOXIDANT ASSAYS
SMALL GTPASE / G-PROTEIN SIGNALING KINASE ASSAYS REPORTER CELLS AND REAGENTS EPITOPE TAGS PROTEIN ANALYSIS TOOLS
CHOLESTEROL/CHOLESTERYL ESTER ASSAYS APOLIPOPROTEIN ASSAYS AND REAGENTS FATTY ACID METABOLISM ASSAYS SERUM PROTEIN ASSAYS RENAL FUNCTION ASSAYS ALCOHOL ASSAYS
VIRUS CORE ANTIGEN ASSAYS TOXIN ASSAYS
ORDERING INFORMATION & SUPPORT
4
Worldwide Technical Support Placing an Order within the U.S.
Placing an Order outside the U.S.
Our Customer Service representatives are available Monday through Friday from 8 am to 5 pm Pacific Time. Most orders received by 2 pm Pacific Time will be shipped the same day. We will notify you immediately of any backordered item. We accept VISA®, MasterCard® and Ameri-can Express® cards. Net 30 day terms may be offered upon credit approval.
Phone 1 858 271 6500 1 888 CBL 0505 (Toll-Free) Fax 1 858 271 6514 E-mail [email protected] Online www.cellbiolabs.com Mail Attn: Customer Service 7758 Arjons Drive San Diego, CA 92126
Pricing
Kit Components
Do you have questions about a particular product before you buy? Do you need help with a protocol? Our Technical Service Scientists have been directly involved in the development and testing of our products, so you get the bene-fit of their hands-on experience.
Phone 1 858 271 6500 1 888 CBL 0505 (US Toll-Free) Fax 1 858 271 6514 E-mail [email protected]
Current U.S. prices are available online at www.cellbiolabs.com, or you may request a separate price list by e-mail. Prices are sub-ject to change without notice. For pricing outside the U.S. please contact your local distributor, which can be found on the inside back cover of this catalog.
Because our kits are QC tested by lot, indi-vidual kit components are generally not available for purchase separately. However, certain components may be available in bulk quantities on a custom basis. For more information please inquire by sending a message to [email protected].
Please see our Worldwide Distributors section on the inside back cover of this cata-log. We have a network of global distributors serving life science researchers in over 70 countries. If you don’t see your country listed, please contact our U.S. office.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
6
CytoSelect™ 96-Well Cell Transformation Assay—Traditional Soft Agar Colony Formation
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation)
Fluorometric 1 Plate* CBA-130
5 Plates* CBA-130-5
Our CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation) is suitable for measur-ing malignant transformation where no downstream analysis is required. Transformed cells cannot be re-covered; however, no manual cell counting is re-quired. With this assay, cells are incubated in a semisolid agar medium for 6-8 days, then solubilized, lysed and detected using CyQuant® GR dye in a fluorometric plate reader.
Fast Results: 6-8 days vs. 21 days Plate Reader Convenience: Eliminates manual
counting Versatile Format: Designed for 96-well through-
put, but can be adapted for 48, 24, 12 or 6-well
CELL-BASED ASSAYS Colony Formation Assays
Transformation of normal cells into neoplastic cells results in a population capable of pro-liferating independently of internal and external signals that normally restrain growth. The soft agar colony formation assay has traditionally been used to monitor anchorage-independent growth, employing 3-4 weeks of cell growth followed by manual cell counting.
We have advanced the soft agar assay to eliminate tedious manual cell counting, allow high-throughput drug screening, and enable recovery of transformed cells for downstream analysis. These advances have also allowed us to develop a unique kit for the separation of clonogenic cancer cells from normal cells in heterogeneous solid tumors.
Tumor Cell / Soft Agar Assays
Cell Transformation Assay Principle.
Recent Product Citations 1. Eisfeld, A. et al. (2014). NRAS isoforms differentially affect
downstream pathways, cell growth, and cell transformation. PNAS 111:4179-4184.
2. Gong, J. et al. (2014). Combined targeting of STAT3/NFkB/COX-2/EP4 for effective management of pancreatic cancer. Clin. Cancer Res. 20:1259-1273.
3. Gupta, P. et al. (2013). Ocrasin targets the JNK-NFkB axis to sensitize glioma cells to TNF-alpha-induced apoptosis. Car-cinogenesis 34:388-396.
4. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103.
5. Rai, V. et al. (2012). Lysophosphatidic acid targets vascular and oncogenic pathways via RAGE signaling. J. Exp. Med. 209:2339-2350.
6. Ghantous, A. et al. (2012). Inhibition of tumor promotion by parthenolide: epigenetic modulation of p21. Cancer Prev. Res. 5:1298-1309.
7. Dennis, M. et al. (2012). Snail controls the mesenchymal phe-notype and drives erlotinib resistance in oral epithelial and head and neck squamous cell carcinoma cells. Carcinogenesis 147:726-732.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively.
CytoSelect™ 96-Well Cell Transformation Assays—Advanced Soft Agar with Post-Incubation Cell Recovery The CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible) provides a robust system for screening oncogenes and cell transformation in-hibitors. Transformed cells may be recovered for fur-ther downstream analysis following colony formation.
Faster Results: 6-8 days vs. 21 days Cell Recovery: Transformed cells remain viable
for further analysis Plate Reader Convenience: Eliminates manual
counting of cells Versatile Format: Designed for 96-well through-
put, but can be adapted for 48, 24, 12 or 6-well
Cell Transformation Assay Principle. Cell colonies form after a 6-8 day incubation with agar matrix. Transformed cells can then be either lysed and detected with a fluorescent dye or recovered and re-plated.
*Each kit provides sufficient reagent quantities to perform 96, 48, 24, 12, or 6 tests in a 96, 48, 24, 12, or 6-well plate, respectively. **The 384-well kit does not allow for cell recovery due to small well size. ***Each kit provides sufficient reagents for one or five 384-well plates respectively.
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible)
Colorimetric 1 Plate* CBA-135
5 Plates* CBA-135-5
Fluorometric 1 Plate* CBA-140
5 Plates* CBA-140-5
CytoSelect™ 384-Well Cell Transformation Assay** 1 Plate*** CBA-145
5 Plates*** CBA-145-5 Fluorometric
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CELL-BASED ASSAYS Colony Formation Assays
Recent Product Citations 1. Singh, R. et al. (2013). Increasing the complexity of chromatin:
functionally distinct roles for replication-dependent histone H2A isoforms in cell proliferation and carcinogenesis. Nucleic Acids Res. 10.1093/nar/gkt736. (CBA-135)
2. Shukla, A. et al. (2013). Extracellular signal-regulated kinase 5: a potential therapeutic target for malignant mesotheliomas. Clin. Cancer Res. 19:2071-2083. (CBA-135)
3. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote im-mortalization, transformation, and migration of primary human foreskin keratinocytes. J. Virol. 86:12384-12396. (CBA-135)
4. Hong, S.W. et al. (2012). Ring finger protein 149 is an E3 ubiq-uitin ligase active on wild-type v-Raf murine sarcoma viral on-cogene homolog B1 (BRAF). J. Biol. Chem. 287:24017-24025. (CBA-135)
5. Lee, H.J. et al. (2012). Cheokine (C-X-C motif) ligand 12 is associated with gallbladder carcinoma progression and is a novel independent poor prognostic factor. Clin. Cancer Res. 18:3270-3280. (CBA-135)
6. Chapeau, E.A. et al. (2012). Ecotropic viral integration site 1 (EVI1) regulates multiple cellular processes important for can-cer and is a synergistic partner for FOS proteinin invasive tu-mors. PNAS 109:2168-2173. (CBA-135)
7. Lim, S.K. et al. (2011). Tyrosine phosphorylation of transcrip-tional coactivator WW-domain binding protein 2 regulates es-trogen receptor alpha function in breast cancer via the Wnt pathway. FASEB J. 25:3004-3018. (CBA-135)
8. Mathew, B. et al. (2011). The novel role of the mu opioid recep-tor in lung cancer progression: A laboratory investigation. Anesth. Analg. 112:558-567. (CBA-135)
9. Hirata, H. et al. (2010). Role of secreted Frizzled-related pro-tein3 in human renal cell carcinoma. Cancer Res. 70:1896-1905. (CBA-135)
10. Hirata, H. et al. (2009). Wnt antagonist gene DKK2 is epigen-etically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways. Clin. Cancer Res. 15:5678-5687. (CBA-135)
www.cellbiolabs.com [email protected]
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay Colorimetric 96 Assays CBA-150
5 x 96 Assays CBA-150-5
CytoSelect™ 96-Well In Vitro Tumor Sensitivity Assay
Inhibition of HeLa Cell Anchorage-Independent Growth by Taxol. HeLa cells were cultured for 7 days in the absence (top) or presence (bottom) of 1 nM Taxol according to the assay protocol.
The CytoSelect™ In Vitro Tumor Sensitivity Assay provides a stringent, anchorage-independent model for chemosensitivity testing and possible anticancer drug screening. The assay uses a soft agar matrix to promote the colony formation of neoplastic cells in about a week. Cells are quantified using a standard ELISA plate reader.
Fast Results: 6-8 days In Vivo Simulation: Resembles a three-
dimensional cell environment Plate Reader Convenience: Eliminates manual
counting
Tumor Sensitivity Assay Principle.
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CELL-BASED ASSAYS Colony Formation Assays
Recent Product Citations 1. Bard-Chapeau, E. et al. (2013). EVI1 oncoprotein interacts with
a large and complex network of proteins and integrates signals through protein phosphorylation. PNAS 110:E2885-E2894.
2. Takezawa, K. et al. (2012). HER2 amplification: a potential mechanism of acquired resistance to EGFR inhibition in EGFR-mutant lung cancers that lack the second-site EGFRT790M mutation. Cancer Discovery 2:922-933.
3. Li,C. et al. (2012). The root bark of Paeonia moutan is a poten-tial anticancer agent in human oral squamous cell carcinoma cells. Anticancer Res. 32:2625-2630.
4. Itamochi, H. et al. (2011). Inhibiting the mTOR pathway syner-gistically enhances cytotoxicity in ovarian cancer cells induced by etoposide through upregulation of c-Jun. Clin. Cancer Res. 17:4742-4750.
5. Kang, D.W. et al. (2010). Phospholipase D1 drives a positive feedback loop to reinforce the Wnt/ß-catenin/TCF signaling axis. Cancer Res. 70:4233-4242.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ Clonogenic Tumor Cell Isolation Kit
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Product Name Size Catalog Number
CytoSelect™ Clonogenic Tumor Cell Isolation Kit 5 Preps CBA-155
25 Preps CBA-155-5
Clonogenic Colony Formation, Isolation and Re-plating. A: Clonogenic colony formation (red arrows) and single cells (black arrows) after 7 day incubation. B: Isolation of clonogenic colonies from single cells. C: Re-plated clonogenic colonies after 3 days (no trypsinization). D: Re-plated clonogenic colonies 1 day after trypsinization.
Clean separation of clonogenic tumor cells from nor-mal cells is critical for proper analysis of disease state progression. Due to the heterogeneity of many tu-mors, however, isolation of homogenous tumor cell populations can be difficult. The CytoSelect™ Clonogenic Tumor Cell Isolation Kit uses a proprietary semisolid agar medium to facilitate colony formation by cells from solid tumors. Colonies are grown in either a 6-well plate or a 35 mm dish. The colonies are then isolated away from single cells by size filtration.
Efficient: Easily eliminates single cells from clonogenic tumor cell population
Versatile: In addition to solid tumors, has potential use in isolating tumor stem cells
Clonogenic Tumor Cell Isolation Procedure.
CELL-BASED ASSAYS Colony Formation Assays
www.cellbiolabs.com [email protected]
CytoSelect™ Leukocyte Endothelium Adhesion Assays
Product Name Detection Size Catalog Number
Fluorometric 96 Assays CBA-210
Fluorometric 96 Assays CBA-211
CytoSelect™ Tumor-Endothelium Adhesion Assay Fluorometric 96 Assays CBA-215
CytoSelect™ Leukocyte-Endothelium Adhesion Assay
CytoSelect™ Leukocyte-Epithelium Adhesion Assay
CytoSelect™ Leukocyte-endothelium Adhesion Assay Principle.
The CytoSelect™ Leukocyte Endothelium Adhesion Assays provide a robust system for the quantitative determination of interactions between leukocytes and endothelium. Adherent cells can be quantified on a fluorescence plate reader.
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CELL-BASED ASSAYS Cell Adhesion
Cell Adhesion Assays
Cell adhesion is a complex mechanism involved in a variety of processes including cell mi-gration/invasion, embryogenesis, wound healing and tissue remodeling. Cells can adhere to the ECM, forming complexes with cytoskeletal components, or to the endothelium.
Our CytoSelect™ Cell Adhesion Assays quantify adhesion of cells using a microplate reader or fluorometer; no manual cell counting is required.
Recent Product Citations 1. Liu, G. et al. (2012). ICAM-1-activated Src and eNOS signal-
ing increase endothelial cell surface PECAM-1 adhesivity and neutrophil transmigration. Blood 120:1942-1952. (CBA-210)
2. Fang, Y. et al. (2012). Site-specific microRNA-92a regulation of Kruppel-like factors 4 and 2 in atherosusceptible endothe-lium. Arterioscler. Thromb. Vasc. Biol. 32:979-987. (CBA-210)
3. Curatola, A.M. et al. (2012). Dehydroepiandrosterone (DHEA) inhibition of monocyte binding by vascular endothelium is associated with sialylation of neural cell adhesion of neural cell adhesion molecule. Repr. Sciences 19:86-91. (CBA-210)
4. Wang, Y.L. et al. (2011). Innate immune function of the adher-ens junction protein p120-catenin in endothelial response to endotoxin. J. Immunol. 186:3180-3187. (CBA-210)
5. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of glioblastoma cells. Neuro Oncology 10.1093/neuonc/nos388. (CBA-215)
6. Hernandez, L. et al. (2010). Activation of NF-kB signaling by inhibitor of NF-kB Kinase ß increases aggressiveness of ovar-ian cancer. Cancer Res. 70:4005-4014. (CBA-215)
CytoSelect™ Microfluidic Biochips
Product Name Detection Size Catalog Number
CytoSelect™ 8-Channel ECM Microfluidic Biochips Microscopy 2 Chips CBA-003
10 Chips CBA-003-5
CytoSelect™ 8-Channel Endothelial Microfluidic Biochips Microscopy 2 Chips CBA-004
10 Chips CBA-004-5
CytoSelect™ 8-Channel Microfluidic Biochips provide an environment that closely mimics in vivo shear stresses, resulting in more physiologically relevant cell adhesion data. The Biochips are self-contained units containing 8 channels with inlet/outlet ports at both ends of each channel. After adding cell samples, a syringe pump set to the proper flow rate applies the appropriate shear stress to the channel.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ 48-well Cell Adhesion Assay. Serum starved cells from three different cell lines were allowed to attach to the ECM-coated plate for 1 hr at 100,000 cells/well. Adherent cells were stained according to the assay protocol.
CytoSelect™ ECM Cell Adhesion Assays
Product Name Detection Size Catalog Number
CytoSelect™ 48-Well Cell Adhesion Assay, ECM Array (Contains one row each of Collagen I, Collagen IV, Fibrinogen, Fibronectin, and Laminin)
Colorimetric 48 Assays CBA-070
5 x 48 Assays CBA-070-5
48 Assays CBA-071
5 x 48 Assays CBA-071-5
CytoSelect™ 48-Well Cell Adhesion Assay, Collagen I Colorimetric 48 Assays CBA-052
Fluorometric 48 Assays CBA-053
CytoSelect™ 48-Well Cell Adhesion Assay, Collagen IV Colorimetric 48 Assays CBA-060
Fluorometric 48 Assays CBA-061
CytoSelect™ 48-Well Cell Adhesion Assay, Fibrinogen Colorimetric 48 Assays CBA-058
Fluorometric 48 Assays CBA-059
CytoSelect™ 48-Well Cell Adhesion Assay, Fibronectin Colorimetric 48 Assays CBA-050
Fluorometric 48 Assays CBA-051
CytoSelect™ 48-Well Cell Adhesion Assay, Laminin Colorimetric 48 Assays CBA-056
Fluorometric 48 Assays CBA-057
Fluorometric
The CytoSelect™ ECM Cell Adhesion Assays provide a quantitative method to measure cell adhesion. The 48-well plate is precoated with your choice of substrate. Adherent cells attach, while non-adherent cells are washed away. Adherent cells can be quantified on a standard plate reader or fluorometer.
Quantitative: Measure results in a colorimetric or fluorescence plate reader
Flexible: Uniform substrate layer of your choice of Collagen I, Collagen IV, Fibrinogen, Fibronectin, or Laminin; or choose the ECM array which contains all 5 ECM proteins
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CELL-BASED ASSAYS Cell Adhesion
BSA
Vitronectin
Laminin
Fibronectin
Collagen IV
Collagen I
MDA-231 HT-1080 HEK293
Recent Product Citations 1. Cervera, A.M. et al. (2008). Cells silenced for SDHB expres-
sion display characteristic features of the tumor phenotype. Cancer Res. 68:4058-4067. (CBA-050 and CBA-070)
2. Lee, J. et al. (2013). Selective inhibition of prostaglandin E2 receptors EP2 and EP4 inhibits adhesion of human endometri-otic epithelial and stromal cells through suppression of integrin-mediated mechanisms. Biol. Reprod. 88:77. (CBA-057)
3. Miao, H. et al. (2008). Gene expression and functional studies of the optic nerve head astrocyte transcriptome from normal African Americans and Caucasian Americans donors. PLoS One 3(8):E2847. (CBA-060)
4. Tanoury, Z. et al. (2014). Genes involved in cell adhesion and signaling: a new repertoire of retinoic acid receptor target genes in mouse embryonic fibroblasts. J. Cell Sci. 127:521-533. (CBA-070)
5. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103. (CBA-070)
6. Mei, J. et al. (2012). Inhibition of IDO1 suppresses cyclooxy-genase-2 and matrix metalloporeinase-9 expression and de-creases proliferation, adhesion and invasion of endometrial stromal cells. Mol. Human Reprod. 10.1093/molehr/gas021. (CBA-070)
7. Kandalam, V. et al. (2011). Lack of tissue inhibitor of metallo-proteinases 2 leads to exacerbated left ventricular dysfunction and adverse extracellular matrix remodeling in response to biomechanical stress. Circulation 124:2094-2105. (CBA-070)
8. Tsukamoto, H. et al. (2010). Evaluation of anticancer activities of benzo[c]phenanthridine alkaloid sanguinarine in oral squamous cell carcinoma cell line. Anticancer Res. 31:2841-2846. (CBA-070)
9. Kim, S.W. et al. (2013). Cardiac stem cells with electrical stimu-lation improve ischaemic heart function through regulation of connective tissue growth factor and miR-378. Cardiovasc. Res. 10.1093/cvr/cvt192. (CBA-071)
www.cellbiolabs.com [email protected]
CELL-BASED ASSAYS Cell Migration / Invasion
12 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Cell Migration & Invasion Assays
Cell migration and invasion are highly integrated, multi-step processes and play important roles in the progression of various diseases including cancer, atherosclerosis and arthritis.
Our cell migration assays are provided in two formats: 2-Dimensional Gap Closure and Boyden Chamber. Each format has its own advantages and applications. Use the informa-tion below to help choose the best format for your cell migration experimental goals. Cell invasion assays are provided in the Boyden Chamber format.
Cell Migration Format Selection Guide
2D Gap Closure Assays
(p. 13) Boyden Chamber Assays
(p. 14-19)
Type of Analysis Qualitative or Quantitative Quantitative
Detection Time Endpoint or Real Time Endpoint
Detection Method Microscopy Plate Reader
Chemoattractant Gradient No Yes
Relative Sensitivity Good Fair
Adaptability to Automation Good Poor
Cell Compatibility Universal Choose pore size based on cell size
Example of Boyden Chamber Assay Principle.
Assay Principle for the Radius™ 2D Gap Closure Assays.
CELL-BASED ASSAYS Cell Migration / Invasion
13 www.cellbiolabs.com [email protected]
Radius™ Cell Migration Assays (2D Gap Closure)
Example Results using 2D Gap Closure Assay.
Product Name Detection Size Catalog Number
Radius™ 24-Well Cell Migration Assay Microscopy 24 Assays CBA-125
5 x 24 Assays CBA-125-5
Radius™ 24-Well Cell Migration Assay (Collagen I Coated) Microscopy 24 Assays CBA-125-COL
Radius™ 24-Well Cell Migration Assay (Fibronectin Coated) Microscopy 24 Assays CBA-125-FN
Radius™ 24-Well Cell Migration Assay (Laminin Coated) Microscopy 24 Assays CBA-125-LN
Radius™ 24-Well Cell Migration Assay (ECM Array Coated) Microscopy 24 Assays CBA-125-ECM
Radius™ 96-Well Cell Migration Assay Microscopy 96 Assays CBA-126
5 x 96 Assays CBA-126-5
Radius™ 384-Well Cell Migration Assay 384 Assays CBA-127
5 x 384 Assays CBA-127-5 Microscopy
Radius™ Cell Migration Assays provide a unique al-ternative to the traditional Boyden Chamber migration assay. Radius assays allow you to measure cell mi-gration at endpoint or in real time, and are ideal for time course migration studies. Radius™ Cell Migration Assays use a cell culture plate containing a proprietary, carefully-defined bio-compatible hydrogel (Radius™ gel) spot centralized at the bottom of each well. Cells seeded in the well will attach everywhere except on the Radius gel spot, creating a cell-free zone. Once cells attach, the Ra-dius gel is removed and migration of cells across the cell-free zone begins. The gel removal step allows synchronization of a zero time point to facilitate well-to-well comparisons. With Radius™ Cell Migration Assays, there are no cell culture inserts; so you don’t need to worry about which pore size to choose for your cell type. Any ad-herent cell may be used in the assay. Radius assays are supplied in 24-well, 96-well and 384-well formats. In addition, the 24-well assays are provided with your choice of coatings for proper cell attachment: Uncoated Collagen I-coated Fibronectin-coated Laminin-coated ECM Array with 6 wells of each of the above
(uncoated, Collagen I, Fibronectin, Laminin); ideal if you are unsure which ECM protein may provide the best cell attachment
Recent Product Citations 1. Sun, J. et al. (2013). Targeting the metastasis suppressor,
NDRG1, using novel iron chelators: regulation of stress fiber-mediated tumor cell migration via modulation of the ROCK1/pMLC2 signaling pathway. Mol. Pharmacol. 83:454-469. (CBA-125)
2. Apostolos, K. et al. (2013). Increased susceptibility of melanin-concentrating hormone-deficient mice to infection with Salmonella enterica serovar Typhimurium. Infect. Immun. 81:166-172. (CBA-125)
3. Smith, K. et al. (2012). Human family with sequence similarity 60 member A (FAM60A) protein: a new subunit of the Sin3 deacety-lase complex. Mol. Cell Proteomics 11:1815-1828. (CBA-125)
4. Young, S. et al. (2012). Rapid protein kinase D1 signaling pro-motes migration of intestinal epithelial cells. Am. J. Physiol. Gas-trointest. Liver Physiol. 303:G356-G366. (CBA-125)
5. Larive, R.M. et al. (2012). The Ras-like protein R-Ras2/TC21 is important for proper mammary gland development. Mol. Biol. Cell 23:2373-2387. (CBA-125)
6. Wong, B. et al. (2013). Adrenomedullin enhances invasion of hu-man extravillous cytotrophoblast-derived cell lines by regulation of urokinase plasminogen activator expression and S-nitrosylation. Biol. Reprod. 88:34. (CBA-126)
7. Ichikawa, A. et al. (2013). CXCL10-CXCR3 enhances the develop-ment of neutrophil-mediated fulminant lung injury of viral and non-viral origin. Am. J. Respir. Crit. Care Med. 187:65-77. (CBA-126)
8. Coulouarn, C. et al. (2012). Hepatocyte-stellate cell cross-talk in the liver engenders a permissive inflammatory microenvironment that drives progression in hepatocellular carcinoma. Cancer Res. 72:2533-2542. (CBA-126)
9. Alcolea, S. et al. (2012). Interaction between head and neck squamous cell carcinoma cells and fibroblasts in the biosynthesis of PGE2. J. Lipid Res. 53:630-642. (CBA-126)
CELL-BASED ASSAYS Cell Migration / Invasion
14
Boyden Chamber Assay Selection Guide
Assay Definition Cell
Types Pore Size
Insert Coating
Assay Formats
Chemotaxis (p. 15)
Migration of cells toward a chemoattractant
(chemical signal) in the cell’s surrounding environment
Neutrophils Leukocytes
3 µm None 24-Well 96-Well
Lymphocytes Monocytes
Macrophages 5 µm None
24-Well 96-Well
Fibroblasts Endothelial Cells Epithelial Cells
Tumor Cells
8 µm None 24-Well 96-Well
Astrocytes Slow-moving Cells
12 µm None 24-Well
Haptotaxis (p. 16)
Migration of cells along a gradient of cellular adhesion sites or extracellular matrix-
bound chemoattractants
Fibroblasts Endothelial Cells Epithelial Cells
8 µm
Collagen I(bottom)
24-Well
Fibronectin(bottom)
24-Well
Transmigration (p. 17)
Migration of cells through the vascular endothelium toward a
chemoattractant
Leukocytes 3 µm None 24-Well
Tumor Cells 8 µm None 24-Well
Invasion (p. 18-19)
Movement of cells through the 3D extracellular matrix into
neighboring tissues; includes ECM degradation and
proteolysis
Fibroblasts Endothelial Cells Epithelial Cells
Tumor Cells
8 µm
ECM Matrix (top)
24-Well 96-Well
Collagen I (top)
24-Well 96-Well
Laminin I (top)
24-Well 96-Well
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ Cell Migration and Invasion Assays (Boyden Chamber)
The Boyden Chamber has been extensively used and widely published as a tool for measuring cell migra-tion and cell invasion in vitro. Our CytoSelect™ Cell Migration and Invasion Assays use this well-cited method to quantify cell migration and invasion with no manual cell counting required. Migratory or invasive cells are quantified using a colorimetric or fluorometric plate reader. Cell migration may take on various forms and behav-iors depending on the type and location of cells. Such subclasses of cell migration include chemotaxis, hap-totaxis, and transmigration. Use the chart below to compare the various subclasses of cell migration as well as cell invasion, which will help you choose the assay best suited to your experimental goals.
Typical Well Setup for Boyden Chamber Assay.
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CytoSelect™ Cell Migration Assays—Chemotaxis
Product Name Pore Size Detection Size Catalog Number
CytoSelect™ 24-Well Cell Migration Assay
3 µm Fluorometric 12 Assays CBA-103
5 x 12 Assays CBA-103-5
5 µm Fluorometric 12 Assays CBA-102
5 x 12 Assays CBA-102-5
8 µm
Colorimetric 12 Assays CBA-100
5 x 12 Assays CBA-100-5
Fluorometric 12 Assays CBA-101
5 x 12 Assays CBA-101-5
12 µm Colorimetric 12 Assays CBA-107
Fluorometric 12 Assays CBA-108
CytoSelect™ 96-Well Cell Migration Assay
3 µm Fluorometric 96 Assays CBA-104
5 x 96 Assays CBA-104-5
5 µm Fluorometric 96 Assays CBA-105
5 x 96 Assays CBA-105-5
8 µm 96 Assays CBA-106
5 x 96 Assays CBA-106-5 Fluorometric
Migration of Human Fibrosarcoma HT-1080 Cells. Cells were seeded at 30,000 cells per well of a 24-well plate and allowed to migrate toward 10% FBS for 4 hours. Migratory cells were stained (above) and quantified in a fluorescence plate reader (data not shown).
Fast Results: Visualize chemotaxis in less than 6 hours with most cell types
Flexible: Bottoms of membrane inserts are un-coated to allow use with any chemoattractant
Higher Throughput: 96-well format available for fluorescence plate readers
CytoSelect™ Cell Migration Assays are ideal for measuring chemotaxis. The kits utilize polycarbonate membrane inserts in 24-well or 96-well plates. Inserts are available with 4 different pore sizes to accommo-date a variety of cell types.
10% FBS 0% FBS
CELL-BASED ASSAYS Cell Migration / Invasion
Recent Product Citations 1. Al-Wadei, M.H. et al. (2013). Gamma-amino butyric acid (GABA)
prevents the induction of nicotinic receptor-reuglated signaling by chronic ethanol in pancreatic cancer cells and normal duct epithe-lia. Cancer Prev. Res. 6:139-148. (CBA-100)
2. Chen, Z. et al. (2012). The iron chelators Cp44mT and DFO in-hibit TGF-ß-induced epithelial-mesenchymal transition via up-regulation of N-Myc downstream-regulated gene 1 (NDRG1). J. Biol. Chem. 287:17016-17028. (CBA-100)
3. Izhak, L. et al. (2010). Predominant expression of CCL2 at the tumor site of prostate cancer patients directs a selective loss of immunological tolerance to CCL2 that could be amplified in a beneficial manner. J. Immunol. 184:1092-1101. (CBA-101)
4. Kiss, J. et al. (2012). Loss of the oxygen sensor PHD3 enhances the innate immune response to abdominal sepsis. J. Immunol. 189:1955-1965. (CBA-102)
5. Verma, S. et al. (2011). Selenoprotein K knockout mice exhibit deficient calcium flux in immune cells and impaired immune re-sponses. J. Immunol. 186:2127-2137. (CBA-103)
6. Li, X. et al. (2011). Kaposi’s sarcoma-associated Herpesvirus-encoded latency-associated nuclear antigen recduces interleukin-8 expression in endothelial cells and impairs neutrophil chemo-taxis by degrading nuclear p65. J. Virol. 85:8606-8615. (CBA-104)
7. Jun, H.S. et al. (2012). Glucose-6-phosphatase-ß, implicated in a congenital neutropenia syndrome, is essential for macrophage energy homeostasis and functionality. Blood 119:4047-4055. (CBA-105)
8. Rosenblum, S. et al. (2012). Timing of intra-arterial neural stem cell transplantation after hypoxia-ischemia influences cell engraft-ment, survival, and differentiation. Stroke 43:1624-1631. (CBA-106)
9. Aftab, B.T. et al. (2011). Itraconazole inhibits angiogenesis and tumor growth in non-small cell lung cancer. Cancer Res. 71:6764-6772. (CBA-106)
www.cellbiolabs.com [email protected]
CELL-BASED ASSAYS Cell Migration / Invasion
CytoSelect™ Cell Migration Assays—Haptotaxis
Product Name Detection Size Catalog Number
CytoSelect™ 24-Well Cell Haptotaxis Assay, Collagen I-coated Colorimetric 12 Assays CBA-100-COL
Fluorometric 12 Assays CBA-101-COL
Colorimetric 12 Assays CBA-100-FN
Fluorometric 12 Assays CBA-101-FN CytoSelect™ 24-Well Cell Haptotaxis Assay, Fibronectin-coated
CytoSelect™ 24-well Cell Haptotaxis Assay. MDA-231 cells were seeded at 150,000 cells/well and allowed to migrate toward FBS for 4 hrs. Migratory cells, found on the bottom of the migration membrane, were stained according to the assay protocol.
Haptotaxis describes the migration of cells toward a gradient of immobilized extracellular matrix. The CytoSelect™ Cell Haptotaxis Assays are ideal for determining the migratory properties of cells. The kits utilize polycarbonate membrane inserts with an 8 µm pore size in a 24-well plate. The undersides of the inserts are coated with either Collagen or Fibronectin. The 8 µm pore size in the membrane inserts is ideal for epithelial cells, endo-thelial cells, fibroblasts, and other cells of similar size. The membrane serves as a barrier that allows discrimination of migratory cells from non-migratory cells.
Fast Results: Visualize cell haptotaxis in less than 6 hours with most cell types
Convenient: Membrane inserts pre-coated on the underside with either Collagen I or Fibronectin
Versatile: Useful with a variety of cell types including epithelial cells, endothelial cells, and fibroblasts*
*For leukocyte migration a 3 µm pore size is recommended. See our CytoSelect™ Chemotaxis Assays (previous page) or the CytoSelect™ Leukocyte Transmigration Assay (next page).
16
Recent Product Citations 1. Herrera, I. et al. (2013). Matrix metalloproteinase (MMP)-1 in-
duces lung alveolar epithelial cell migration and proliferation, protects from apoptosis, and represses mitochondrial oxygen consumption. J. Biol. Chem. 288:25964-25975. (CBA-100-COL)
2. Niccoli, S. et al. (2012). The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote immor-talization, transformation, and migration of primary human fore-skin keratinocytes. J. Virol. 86:12384-12396. (CBA-110-COL)
3. Kamiya, K. et al. (2007). Protein Kinase C delta activated adhe-sion regulates vascular smooth muscle cell migration. J. Surg. Res. 141:91-96. (CBA-100-COL)
Assay Principle for the CytoSelect™ Cell Haptotaxis Assay.
BSA Collagen I Fibronectin
0% FBS
0.5% FBS
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CELL-BASED ASSAYS Cell Migration / Invasion
17
CytoSelect™ Cell Migration Assays—Transmigration
Product Name Detection Size Catalog Number
CytoSelect™ Leukocyte Transmigration Assay Fluorometric 24 Assays CBA-212
Fluorometric 24 Assays CBA-216 CytoSelect™ Tumor Transendothelial Migration Assay
Pore Size
3 µm
8 µm
Cancer cell transmigration, particularly extravasation, is an important step in cancer metastasis. The CytoSe-lect™ Cell Transmigration Assays provide a robust system for the quantitation of transmigrations and interac-tions between endothelium and cancer cells. Migratory cells are quantified via fluorometer.
Recent Product Citations 1. Fava, G. et al. (2008). Leptin enhances cholangiocarcinoma cell
growth. Cancer Res. 68:6752-6761. (CBA-212) 2. Park, G.B. et al. (2014). The Epstein-Barr Virus causes epithelial
-mesenchymal transition in human corneal epithelial cells via Syk/Src and Akt/ERK signaling pathways. Invest. Ophthalmol. Vis. Sci. 55:1770-1779. (CBA-216)
3. Xu, Z. et al. (2010). Role of pancreatic stellate cells in pancreatic cancer metastasis. Am. J. Pathol. 177:2585-2596. (CBA-216)
Assay Principle for the CytoSelect™ Leukocyte Transmigration Assay.
The Leukocyte Adhesion and Transmigration Cascade.
www.cellbiolabs.com [email protected]
0
40
80
120
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Quantitation of Human Monocytic THP-1. LeukoTracker™ labeled THP-1 cells were titrated in 1X PBS, then lysed with 2X lysis buffer. Fluorescence was quantified as described in the assay protocol.
CytoSelect™ Cell Invasion Assays
Quantitative: Measure results in a colorimet-ric or fluorescence plate reader
Flexible: Uniform protein matrix layer of your choice of basement membrane (from mouse tumor cells), Collagen I, or Laminin I
Versatile: Characterize both the invasive and migratory properties of your cells with a Cell Migration / Invasion Combo Kit (next page)
CytoSelect™ Cell Invasion Assay Principle.
Human Fibrosarcoma HT-1080 Laminin I Cell Invasion. HT-1080 and NIH3T3 (negative control) were seeded at 200,000 cells/well and allowed to invade toward FBS for 24 hrs. Invasive cells on the membrane bottom were stained (top and center) and quantified at OD 560nm after extraction (data not shown).
Tumor cell invasion into surrounding normal tissue contributes to the morbidity of cancers. The CytoSe-lect™ Cell Invasion Assays use precoated inserts to assay invasive properties of tumor cells in 24-well or 96-well plates. The coated layer serves to distinguish invasive cells from non-invasive cells. Plates are pre-coated with either basement membrane matrix (from EHS mouse sarcoma cells), Collagen I or Laminin I.
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CELL-BASED ASSAYS Cell Migration / Invasion
Recent Product Citations 1. Gradilone, S. et al. (2013). HDAC6 inhibition restores ciliary
expression and decreases tumor growth. Cancer Res. 73:2259-2270. (CBA-110)
2. Nam, H. et al. (2013). Antitumor activity of saracatinib (AZD0530), a c-Src/Abl kincase inhibitor, alone or in combina-tion with chemotherapeutic agents in gastric cancer. Mol. Can-cer Ther. 12:16-26. (CBA-110)
3. DiNatale, B. et al. (2012). Ah receptor antagonism represses head and neck tumor cell aggressive phenotype. Mol. Cancer Res. 10:1369-1379. (CBA-110)
4. Citterio, C. et al (2012). The Rho exchange factors Vav2 and Vav3 control a lung metastasis-specific transcriptional program in breast cancer cells. Sci. Signal. 5:ra71. (CBA-110)
5. Nakayama, K. et al. (2013). cAMP-response element-binding protein (CREB) and NFkB transcription factors are activated during prolonged hypoxia and cooperatively regulate the induc-tion of matrix metalloproteinase MMP1. J. Biol. Chem. 288:2584-22595. (CBA-112)
6. Desai, S.D. et al. (2012). ISG15 disrupts cytoskeletal architec-ture and promotes motility in human breast cancer cells. Exp. Biol. Med. 237:38-49. (CBA-112)
7. Ziemann, A. et al. (2013). CRN2 enhances the invasiveness of gliobastoma cells. Neuro Oncology 10.1093/neuonc/nos388. (CBA-112-COL)
8. Liu, X. et al. (2013). Antiproliferative, antiinvasive, and proapoptotic activity of folate receptor alpha-targeted liposomal doxorubicin in nonfunctional pituitary adenoma cells. Endocri-nology 154:1414-1423. (CBA-112-COL)
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ Cell Invasion Assays, continued
Product Name Detection Size Catalog Number
CytoSelect™ 24-Well Cell Invasion Assay, Basement Membrane Colorimetric 12 Assays CBA-110
Fluorometric 12 Assays CBA-111
CytoSelect™ 24-Well Cell Invasion Assay, Collagen I Colorimetric 12 Assays CBA-110-COL
Fluorometric 12 Assays CBA-111-COL
Colorimetric 12 Assays CBA-110-LN
Fluorometric 12 Assays CBA-111-LN
CytoSelect™ 96-Well Cell Invasion Assay, Basement Membrane Fluorometric 96 Assays CBA-112
CytoSelect™ 96-Well Cell Invasion Assay, Collagen I Fluorometric 96 Assays CBA-112-COL
CytoSelect™ 96-Well Cell Invasion Assay, Laminin I Fluorometric 96 Assays CBA-112-LN
CytoSelect™ 24-Well Cell Invasion Assay, Laminin I
CytoSelect™ Cell Migration / Invasion Assay Combo Kits
Effects of Cytochalasin D on Invading Cells using the CytoSelect™ 24-well Cell Invasion Assay (CBA-110). HT-1080 and NIH3T3 cells (negative control) were seeded at 300,000 cells/well and allowed to invade toward 10% FBS for 24 hrs, in the presence or absence of 2 µM Cytochalasin D. Invasive cells, on the bottom of the invasion membrane, were stained (left) and then quantified at OD 560 nm after ex-traction using a standard plate reader (right).
Our CytoSelect™ Cell Migration / Invasion Assay Combo Kits allow you to characterize both the migratory and invasive properties of your cells. Each 24-well combo kit provides sufficient reagents to perform 12 migration plus 12 invasion assays, while the 96-well combo kit allows you to perform 96 migration plus 96 invasion assays. The invasion plate provided contains basement membrane-coated inserts.
19
NIH3T3 HT-1080
0
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NIH3T3 HT-1080 HT-1080 +Cytochalasin D
OD
560
nm
CELL-BASED ASSAYS Cell Migration / Invasion
Recent Product Citations 1. Majid, S. et al. (2013). miRNA-34b inhibits prostate cancer through demethylation, active chromatin modification, and AKR pathways. Clin.
Cancer Res. 19:73-84. (CBA-100-C) 2. Shin, S.Y. et al. (2012). Transcriptional regulation of the interleukin-11 gene by oncogenic Ras. Carcinogenesis 10.1093/carcin/bgs297.
(CBA-100-C) 3. Gobeil, S. et al. (2008). A genome-wide shRNA screen identifies GAS1 as a novel melanoma metastasis suppressor gene. Genes Dev.
22(21):2932-2940. (CBA-101-C) 4. Axlund, S.D. et al. (2010). HOXC8 inhibits androgen receptor signaling in human prostate cancer cells by inhibiting SRC-3 recruitment to
direct androgen target genes. Mol. Cancer Res. 8:1643-1655. (CBA-106-C)
Product Name Pore Size Detection Size Catalog Number
CytoSelect™ 24-Well Cell Migration / Invasion Combo Kit Colorimetric 2 x 12 Assays CBA-100-C
Fluorometric 2 x 12 Assays CBA-101-C
CytoSelect™ 96-Well Cell Migration / Invasion Combo Kit 8 µm Fluorometric 2 x 96 Assays CBA-106-C
8 µm
www.cellbiolabs.com [email protected]
20
CELL-BASED ASSAYS Cell Migration / Invasion
CytoSelect™ 24-Well Wound Healing / Cell Migration Assay
Highly Accurate: More consistent results well-to-well compared to homemade scratch assays
Versatile: Measure cell migration, cell prolifera-tion, and wound closure
Inert Material: No residues from inserts to impede cell migration or proliferation
Compared to traditional scratch assays, our CytoSe-lect™ 24-Well Wound Healing Assay provides a more consistent method to measure cell migration across a “wound field” gap in vitro. Proprietary treated inserts generate a consistently defined 0.9mm gap between the cells. Cells can then be treated and monitored for proliferation or migration across the wound field by imaging samples at fixed time points or time-lapse microscopy.
Product Name Detection Size Catalog Number
CytoSelect™ 24-Well Wound Healing Assay 24 Assays CBA-120
5 x 24 Assays CBA-120-5 Microscopy
Wound Closure of STO Cells. STO cells (mouse MEF) were cul-tured in the provided plate with inserts in place for 24 hours until a monolayer formed. Inserts were then removed to begin the assay. Cells were monitored at various time points and stained according to the assay protocol. CytoSelect™ 24-well Wound Healing Assay Principle.
0%
50%
100%
Percent Wound Closure
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citations 1. Cui, X.G. et al. (2013). Response gene to complement 32 defi-
ciency causes impaired placental angiogenesis in mice. Cardio-vasc. Res. 10.1093/cvr/cvt121.
2. Gutschner, T. et al. (2013). The noncoding RNA MALAT1 is a critical regulator of the metastasis phenotype of lung cancer cells. Cancer Res. 73:1180-1189.
3. Xing, C. et al. (2013). Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma. Mol. Cancer Ther. 12:94-103.
4. Mao, R. et al. (2012). Circadian gating of epithelial-to-mesenchymal transition in breast cancer cells via melatonin-regulation of GSK3ß. Mol Endrocrinol. 26:1808-1820.
5. Hirata, H. et al. (2012). MicroRNA-1826 targets VEGFC, beta-catenin (CTNNB1) and MEK1 (MAP2K1) in human bladder can-cer. Carcinogenesis 33:41-48.
21
CELL-BASED ASSAYS Cell Co-Culture
www.cellbiolabs.com [email protected]
Product Name Detection Size Catalog Number
CytoSelect™ 24-Well Cell Co-Culture System Microscopy 24 Assays CBA-160
CytoSelect™ 24-Well Cell Co-Culture System
Traditional methods of co-culture usually involve one of the following methods: 1. One cell type is cultured to form a monolayer,
followed by seeding of a second cell type directly over the monolayer. This is a common method when feeder cells are used to maintain stem cells in an undifferentiated state. However, it is not useful when studying the effects of one cell on the other because the first cell is obscured from view by the second.
2. A Boyden Chamber (see page 12 for details) is used to culture one cell type above the mem-brane and a second cell type below the mem-brane. This system allows a separation between the two cells, but does not allow for direct cell-to-cell contact which may reduce its efficacy for cer-tain applications.
The CytoSelect™ Cell Co-Culture System provides a unique platform for direct contact between two cell types in one well. A proprietary molded plastic insert creates a cell-free zone in the center of a 24-well cell culture-treated plate. The first cell type is seeded in the area around the insert. Once the cells form a monolayer, the insert is removed and the second cell is seeded.
The culture of two cell lines together is advantageous for studying a variety of applications: Cell-cell interactions Cell activation Cellular differentiation Maintaining stem cell pluripotency Various effects of secreted factors from one cell type on a second cell type
Protocol for the CytoSelect™ 24-Well Co-Culture System. Cell type #1 is seeded with the Co-Culture insert in place. After cells form a monolayer, the insert is removed and cell type #2 is seeded.
CELL-BASED ASSAYS Cell Health
22
CytoSelect™ Cell Viability and Cytotoxicity Assay (Live/Dead)
Product Name Detection Size Catalog Number
CytoSelect™ Cell Viability and Cytotoxicity Assay Kit Colorimetric / Fluorometric
96 Assays CBA-240
Cell viability characteristics include cellular metabolic activity and cell membrane integrity. Our CytoSe-lect™ Cell Viability and Cytotoxicity Assay provides both a colorimetric and fluorometric format for moni-toring cell viability via metabolic activity. Live cells are detected with MTT (colorimetric detection) or Calcein AM (fluorometric); dead cells are detected with EthD-1 reagent (fluorometric). All 3 detection reagents are included, as well as Saponin, a cell death initiator. Cells may be treated with compounds or agents that affect cell viability. This kit is suitable for eukaryotic cells, not bacteria or yeast.
Viability of Human Foreskin Fibroblasts. BJ-TERT cells were seeded at 50,000 cells/well and allowed to culture for 24 hours. Cells were then treated with and without Saponin. All cells were then stained with Calcein AM and EthD-1. Top: Cells without Saponin treatment. Bottom: Cells with Saponin treatment. Left: Calcein AM staining. Right: EthD-1 staining.
Versatile: Detect by microscopy, colorimetric or fluorescence plate reader, or flow cytometry
Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citation Kim, E.Y. et al. (2012). Sustained activation of N-methyl-D-aspartate receptors in podocytes leads to oxidative stress, mobili-zation of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737.
CytoSelect™ LDH Cytotoxicity Assay
Product Name Detection Size Catalog Number
CytoSelect™ LDH Cytotoxicity Assay Kit Colorimetric 960 Assays CBA-241
Loss of cell membrane integrity is one of the hall-marks of cytotoxicity. Upon cell death, lactate dehy-drogenase (LDH) is released from the cytoplasm through the damaged membrane. Our CytoSelect™ LDH Cytotoxicity Assay provides a convenient plate-based method for testing cytotoxicity based on LDH release. In this assay, cells are cul-tured in a 96-well plate with and without the com-pound to be tested. LDH released into the media from cells converts a lactate substrate to pyruvate and generates NADH. In the presence of NADH, the col-orimetric dye WST-1 is converted to a formazan that generates an orange color. Detection is performed in a standard colorimetric plate reader.
LDH Release from HEK 293 Cells. 20,000 cells/well were cultured for 24 hours. After adding various concentrations of Triton X-100, the LDH Cytotoxicity Assay Reagent was added followed by a 30 minute incubation at 37ºC and 5% CO2.
CytoSelect™ Anoikis Assays
Product Name Detection Size Catalog Number
CytoSelect™ 24-Well Anoikis Assay Colorimetric / Fluorometric
24 Assays CBA-080
CytoSelect™ 96-Well Anoikis Assay Colorimetric / Fluorometric
96 Assays CBA-081
Anoikis of Human Fibroblast BJ-TERT Cells. 50,000 cells/well were seeded in a control plate (left) and a Poly-HEMA coated plate (right) and cultured for 24 hours. Cells on the control plate were stained with Calcein AM. Cells on the Poly-HEMA coated plate were stained with EthD-1.
Anoikis is defined as death of adherent cells due to loss of adhesion to the extracellular matrix. Our Anoikis Assays allow you to quantify and monitor an-chorage-dependent cell death using a precoated plate. Live cells can be viewed under a microscope and quantified on a plate reader by MTT (colorimetric) or Calcein AM (fluorometric), both in-cluded with the kit. Dead cells are detected with an EthD-1 reagent.
Versatile: Detect live and dead cells by micros-copy, fluorescence, or flow cytometry
Quantitative: Measure live and dead cells on a fluorescence plate reader; live cells may also be quantified on a standard microplate reader
Cellular Senescence Assays
Product Name Detection Size Catalog Number
Cellular Senescence Assay Kit (SA -gal Staining) Light Microscopy 50 Assays CBA-230
5 x 50 Assays CBA-230-5
96-Well Cellular Senescence Assay (SA -gal Activity) Fluorometric Plate Reader
120 Assays CBA-231
5 x 120 Assays CBA-231-5
Flow Cytometry / Fluorescence Microscopy
10 Assays CBA-232
5 x 10 Assays CBA-232-5 Quantitative Cellular Senescence Assay (SA -gal)
Senescence Associated (SA) -galactosidase is a common biochemical marker of cellular senescence. Cells expressing such markers have been identified in vivo in tissues. SA ß-Gal catalyzes hydrolysis of X-gal, which produces a blue color in senescent cells. We offer three kit formats to test cellular senescence via SA-ß-galalactosidase activity: Our ß-Gal Staining Kit allows you to visualize senescence by standard light microscope. Our Quantitative Cellular Senescence Assay measures senescence in cells cultured in a 35mm dish by
either flow cytometry or fluorescence microscopy Our 96-Well Cellular Senescence Assay provides a higher throughput assay in a fluorescence plate reader.
Recent Product Citations 1. Lee, H.W. et al. (2013). Tpl2 kinase impacts tumor growth
and metastasis of clear cell renal cell carcinoma. Mol. Can-cer Res. 11:1375-1386. (CBA-080)
2. Sisto, M. et al. (2009). Fibulin-6 expression and anoikis in human salivary gland epithelial cells: implications in Sjogren's syndrome. Int. Immunol. 21:303-311. (CBA-080)
3. Liu, H. et al (2008). Cysteine-rich protein 61 and connective tissue growth factor induce de-adhesion and anoikis of reti-nal pericytes. Endocrinology 149:1666-1677. (CBA-080)
CELL-BASED ASSAYS Cell Health
23 www.cellbiolabs.com [email protected]
Recent Product Citation Malhotra, D. et al. (2010). Global mapping of binding sites for Nrf2 identifies novel targets in cell survival response through ChIP-Seq profil-ing and network analysis. Nucleic Acid Res. 10.1093/nar/gkq212. (CBA-231)
CELL-BASED ASSAYS Cell Health
24 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ MTT Cell Proliferation Assay
Product Name Detection Size Catalog Number
CytoSelect™ MTT Cell Proliferation Assay Colorimetric 960 Assays CBA-252
Cell proliferation is easily measured by the addition of a variety of dyes that produce a visible color that can be correlated with the number of cells. Our CytoSelect™ MTT Cell Proliferation Assay provides a simple method to measure proliferation of cells. The cell-permeable MTT dye is added directly to cultured cells followed by a de-tergent solution. Quantitation is performed using a standard microplate reader at 540-570 nm.
CytoSelect™ WST-1 Cell Proliferation Assay Reagent
Our CytoSelect™ WST-1 Cell Proliferation Assay provides a similar method to our MTT Cell Proliferation Assay, but with a single reagent format that does not require a detergent solubilization step. Quantitation is performed using a standard microplate reader at 450 nm.
Product Name Detection Size Catalog Number
CytoSelect™ Cell Proliferation Assay Reagent, Fluorometric Fluorometric 960 Assays CBA-250
CytoSelect™ Fluorometric Cell Proliferation Assay Reagent
Product Name Detection Size Catalog Number
CytoSelect™ WST-1 Cell Proliferation Assay Reagent Colorimetric 960 Assays CBA-253
Human HEK 293 Cell Density. Cells were seeded at various den-sities in triplicate and allowed to culture for 24 hours. Cells were then treated with the CytoSelect™ Cell Proliferation Assay Reagent for 6 hours at 37ºC and 5% CO2.
Cell proliferation is easily measured by the addition of a variety of dyes that can be correlated with the num-ber of cells. Various dyes producing a visible color are available to measure proliferation rates, but fluorometric dyes are often more sensitive and may be a superior choice for researchers with access to a fluorescence-based microplate reader. Our CytoSelect™ Cell Proliferation Assay Reagent (Fluorometric) provides a simple, single reagent method to measure proliferation of cells. The fluoro-metric dye is added directly to cultured cells. Upon entering metabolically active live cells, the non-fluorescent dye is converted to a bright red fluores-cent form. Quantitation is performed using a fluores-cence plate reader with excitation at 560 nm and emission at 590-600 nm. This reagent is versatile and can be used with a wide variety of cell types including cultured mammalian and piscine cells, bacteria, yeast, fungi, and protozoa.
CELL-BASED ASSAYS Cell Health
25 www.cellbiolabs.com [email protected]
CytoSelect™ BrdU Cell Proliferation ELISA Kit
Product Name Detection Size Catalog Number
CytoSelect™ BrdU Cell Proliferation ELISA Kit Colorimetric 96 Assays CBA-251
Assay Principle for the CytoSelect™ BrdU Cell Proliferation ELISA Kit.
BrdU is a thymidine analog that can incorporate into newly synthesized DNA strands of actively proliferating cells. Our CytoSelect™ BrdU Cell Proliferation ELISA Kit provides a convenient plate-based method to measure this incorporation. Once the BrdU is incorporated into the DNA, cells are fixed and DNA is denatured. Incorpo-rated BrdU can be quantified in the denatured DNA by an anti-BrdU antibody. The absorbance readings at 450 nm can be directly correlated to cell proliferation.
CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit
Product Name Detection Size Catalog Number
CytoSelect™ Proliferating Cell Nuclear Antigen (PCNA) ELISA Kit Colorimetric 96 Assays CBA-254
PCNA Detection in HeLa Whole Cell Lysates. Whole cell lysates were prepared in a RIPA lysis buffer. Protein concentrations were determined by BCA protein assay.
Proliferating Cell Nuclear Antigen (PCNA) acts as a processivity factor for DNA polymerase by associat-ing with various proteins involved in DNA replication. It is also associated with chromatin remodeling and cell cycle control, and it is often used as a marker of cell proliferation. Our CytoSelect™ PCNA ELISA Kit provides a con-venient plate-based method to quantify PCNA levels in nuclear or whole cell extracts.
Sensitive: Detect PCNA as low as 12.5 ng/mL Versatile: Measure PCNA levels from human,
mouse, or rat whole cell lysates or nuclear extracts Quantitative: Measure results in a colorimetric
plate reader against a provided PCNA standard
CELL-BASED ASSAYS Cell Hypoxia
26 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
HIF-1 Alpha DNA Binding Activity Assay Kit
Detection Specificity of HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM deferoxamine mesylate (DFO) for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit (#AKR-171). 100 pmol of non-biotinylated wild type or mutated HRE double stranded com-petitor oligos were added to the Complete DNA Binding Buffer just prior to inclusion in the assay.
Product Name Detection Size Catalog Number
HIF-1 Alpha DNA Binding Activity Assay Kit Colorimetric 96 Assays CBA-282
Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it can also be a symptom of pathological conditions and is often used as a marker for tumor cells. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hy-poxia-responsive genes such as erythropoietin and VEGF. During hypoxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes transcriptionally active. It then binds tran-scriptional coactivators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit de-tects activated HIF-1 in an ELISA format. Active HIF-1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRP-conjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat protein samples.
Product Name Detection Size Catalog Number
HIF-1 Alpha Sandwich ELISA Kit Colorimetric 96 Assays CBA-280
HIF-1 Alpha Cell Based ELISA Kit Chemiluminescent 96 Assays CBA-281
HIF-1 Alpha ELISA Kits
Detection of Nuclear HIF-1 Alpha with the HIF-1 Alpha Sand-wich ELISA Kit. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) nuclear extracts according to the Assay Protocol.
Our HIF-1 Alpha ELISA Kits provide a convenient method for detection and quantitation of human, mouse, or rat HIF-1 Alpha in cells or tissues. Two ELISA kit formats are available: The HIF-1 Alpha Sandwich ELISA Kit detects HIF
-1 Alpha in any protein sample including tissue homogenates, whole cell lysates, or nuclear ex-tracts. Samples are added to an anti-HIF-1 Alpha antibody coated plate. Quantitation of unknown samples is performed by comparison of the OD values to those of a known standard.
The HIF-1 Alpha Cell Based ELISA Kit allows the detection of HIF-1 Alpha levels in intact cells. Cells are seeded in a tissue culture treated plate suitable for reading in a 96-well plate-based lumi-nometer. Cells are fixed and permeabilized to allow detection with the anti-HIF-1 antibody. De-tection is performed by chemiluminescence.
CELL-BASED ASSAYS Adipogenesis
27 www.cellbiolabs.com [email protected]
CytoSelect™ 96-well Adipogenesis Assay Kit
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well Adipogenesis Assay Kit Colorimetric / Fluorometric
200 Assays CBA-290
Staining of 3T3-L1 Cells with Oil Red O. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded overnight in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Oil Red O colorimetric stain according to the Assay Protocol.
Staining of 3T3-L1 Cells with Nile Red Fluorescent Stain. 20,000 cells/well of preadipocyte 3T3-L1 cells were seeded over-night in a 96-well plate. Cells were uninduced (top) or induced (bottom) for 7 days and stained with Nile Red Fluorescent Stain according to the Assay Protocol.
The ability to regulate the cell cycle and differentiation of adipocytes is important to the understanding of obesity. Adipogenesis is the process in which preadipocytes develop into mature adipocytes in a multistep process that requires the sequential activation of numerous transcription factors. The 3T3-L1 cell line is the best character-ized model for adipogenesis in vitro. 3T3-L1 cells display a fibroblast-like phenotype when grown under normal conditions. However, when treated with a combination of IBMX, insulin, and dexamethasone, these cells un-dergo terminal differentiation resulting in a more rounded phenotype and the formation of intracellular lipid drop-lets. The CytoSelect™ 96-Well Adipogenesis Assay quantitatively measures lipid droplet accumulation in cultured cells of the 3T3-L1 model. Quantitation is performed either in a standard colorimetric plate reader with Oil Red O stain, or in a fluorescence plate reader with Nile Red fluorometric stain.
CELL-BASED ASSAYS Phagocytosis
28 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CytoSelect™ 96-Well Phagocytosis Assays
Phagocytosis may be assayed by measuring the en-gulfing of a cell “substrate” such as an erythrocyte(RBC) or Zymosan particle. Traditional phagocytosis assays involve manually counting the engulfed sub-strates under a microscope. This process is tedious and time-consuming, can be somewhat inaccurate, and is not amenable to high throughput. CytoSelect™ 96-Well Phagocytosis Assays are more accurate, high-throughput alternatives to the standard phagocytosis assay. The assays may be adapted for use in 48-well and 24-well plates if desired.
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well Phagocytosis Assay (E. coli) Colorimetric 96 Assays CBA-222
CytoSelect™ 96-Well Phagocytosis Assay (Red Blood Cell) Colorimetric 96 Assays CBA-220
Colorimetric 96 Assays CBA-224
5 x 96 Assays CBA-224-5 CytoSelect™ 96-Well Phagocytosis Assay (Zymosan)
Assay Principle for the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan).
Particle Engulfment with the CytoSelect™ 96-Well Phagocytosis Assay (Zymosan).
Recent Product Citations 1. Lee, J.K. et al. (2010). Regulator of G-protein signaling-10 nega-
tively regulates NF-kB in microglia and neuroprotects dopa-minergic neurons in hemiparkinsonian rats. J. Neurosci. 31:11879-11888. (CBA-220)
2. Winnicka, B. et al. (2010). CD13 is dispensible for normal hema-topoiesis and myeloid cell functions in the mouse. J. Leukoc. Biol. 88(2):347-359. (CBA-220)
3. Hamilton, C.M. et al. (2009). Fasciola hepatica tegumental anti-gen suppresses dendritic cell maturation and function. Infect. Immun. 77:2488-2498. (CBA-220)
4. Dowling, D.J. et al. (2009). Major secretory antigens of the helminth Fasciola hepatica activate a suppressive dendritic cell phenotype that attenuates Th17 cells but fails to activate Th2 immune responses. Infect. Immun. 78:793-801. (CBA-220)
5. Pierce, L.M. et al. (2012). Effect of heavy metal tungsten alloy particles on oxidative product formation and phagocytosis by lung macrophages. Am. J. Respir. Crit. Care Med. 185:A4666. (CBA-224)
6. Polancec, D.S. et al. (2012). Azithromycin drives in vitro GM-CSF/IL-4-induced differentiation of human blood monocytes toward dendritic-like cells with regulatory properties. J. Leukoc. Biol. 91:229.-243. (CBA-224)
Highly Accurate: Eliminates manual counting High Throughput: 96-well plate format Quantitative: Measure OD in a standard microplate
reader Flexible: Choose from 3 substrates: E. coli, Zymo-
san particles, or red blood cells*
*Red blood cells are not provided in the kit. Fresh RBCs should be obtained immediately prior to running the assay. E. coli and Zymosan particles are provided in their respective kits.
CELL-BASED ASSAYS Cell Contraction and Angiogenesis
29 www.cellbiolabs.com [email protected]
Endothelial Tube Formation (In Vitro Angiogenesis) Assay
Product Name Detection Size Catalog Number
Endothelial Tube Formation Assay (In Vitro Angiogenesis) Light Microscopy 50 Assays CBA-200
Recent Product Citations 1. Yu, J.G. et al. (2013). Baroreflex deficiency hampers angiogene-
sis after myocardial infarction via acetylcholine-alpha7-nicotinic ACh receptor in rats. Eur. Heart J. 34:2412-2420.
2. Bid, H. et al. (2013). Dual targeting of the Type 1 insulin-like growth factor receptor and its ligands as an effective antiangio-genic strategy. Clin. Cancer Res. 19:2984-2994.
3. Cai, C. et al. (2012). SIVmac239-Nef downregulates cell surface expression of CXCR4 in tumor cells and inhibits proliferation, migration and angiogenesis. Anticancer Res. 23:2759-2768.
4. Bid, H. et al. (2012). Potent inhibition of angiogenesis by the IGF-1 receptor-targeting antibody SCH717454 is reversed by IGF-2. Mol. Cancer Ther. 11:649-659.
For angiogenesis to occur, endothelial cells must es-cape their stable location and break through the basement membrane. These cells proliferate to form new blood vessels. Our Endothelial Tube Formation Assay provides an easy, robust system to assess angiogenesis in vitro. The assay uses an ECM gel matrix derived from mouse sarcoma cells; this matrix very closely resembles an in vivo basement mem-brane environment.
Product Name Detection Size Catalog Number
Cell Contraction Assay Light Microscopy 24 Assays CBA-201
Collagen-Based Cell Contraction Assay
Assay Principle for the Collagen-Based Cell Contraction Assay.
Recent Product Citations 1. Kotio, K.U. et al. (2011). Implication of microRNAs in atrial natri-
uretic peptide and nitric oxide signaling in vascular smooth mus-cle cells. Am. J. Physiol. Gastrointest. Liver Physiol. 301: C929-C937.
2. Schell, C. et al. (2010). 15-deoxy-delta12-14-prostaglandin-J2 induces hypertrophy and loss of contractility in human testicular peritubular cells: implications for human male fertility. Endocri-nology 151:12571268.
Wound healing is comprised of epithelialization, con-nective tissue deposition, and contraction. The con-traction process is believed to be mediated by spe-cialized fibroblasts (myofibroblasts). 3D collagen gels have been widely used in fibroblast contraction stud-ies. Our Cell Contraction Assay provides a simple system to assess cell contractivity and to screen for cell con-traction mediators. The system uses a 3D collagen matrix to measure changes in the collagen gel size. An optional contraction inhibitor is provided.
HUVEC Tube Formation on ECM Gel. HUVEC cells from a stan-dard tissue culture plate were incubated on an ECM gel. After sev-eral hours tube formation can be visualized under a light micro-scope.
CELL-BASED ASSAYS Autophagy
30
GFP-LC3 Expression Vectors
Product Name Size Catalog Number
pCMV-GFP-LC3 Expression Vector 100 µL CBA-401
pSMPUW-GFP-LC3 Lentiviral Expression Vector 10 µg LTV-801
10 µg RTV-801 pMXs-GFP-LC3 Retroviral Expression Vector
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citations 1. Chen, W. et al. (2012). Andrographolide induces autophagic cell death in human liver cancer cells through cyclophilin D-mediated mito-
chondrial permeability transition pore. Carcinogenesis 10.1093/carcin/bgs264. (CBA-401) 2. Cina, D.P. et al. (2012). Inhibition of MTOR disrupts autophagic flux in podocytes. J. Am. Soc. Nephrol. 23:412-420. (CBA-401) 3. Tu, S.P. et al. (2011). IFN-gamma inhibits gastric carcinogenesis by inducing epithelial cell autophagy and T-cell apoptosis. Cancer Res.
71:4247-4259. (CBA-401)
MAP LC3 is the most published autophagosome marker protein. LC3 associates to the inner and outer limiting membranes of the auto-phagosome. There are two forms of LC3 visible by immunoblot: LC3I which is found in the solu-ble fraction, and LC3II which is found in the membrane fraction. The proportion of LC3II in-creases during autophagy. Our GFP-LC3 expression vectors are conven-ient tools for the study of autophagy. These vec-tors are available in three formats: mammalian, lentiviral, and retroviral expression vectors. Each vector contains a GFP reporter gene. In addi-tion, a GFP control plasmid is provided at no additional charge.
STEM CELL RESEARCH Induced Pluripotent Stem Cells
iPS Cell Reprogramming
Reprogramming of adult cells into induced pluripotent stem cells (iPS) has provided an im-portant new vehicle to facilitate stem cell research. Recent studies have shown that this may be accomplished by the introduction of key genes into somatic cells by transduction with various viral vectors or transfection of plasmids. Retroviral and lentiviral vectors appear to achieve among the highest levels of efficiency of iPS cell generation. We offer an extensive collection of vectors for iPS cell reprogramming.
32
Retroviral Vectors and Packaging Cells for iPS Cell Generation
Our iPS retroviral vectors are constructed from the pMXs vector backbone developed by Dr. Toshio Kitamura at the University of Tokyo.* Each vector contains one of 6 factors shown to help reprogram adult fibroblasts into iPS cells. Both human and mouse genes are available individually or in sets. Separate retroviral vectors are available for p53 shRNA, which has been shown to potentially increase the efficiency of iPS cell generation. Platinum Retroviral Packaging Cells provide an easy way to produce high-titer retroviruses from these stem cell plasmids. For additional information on these cell lines please see p. 64.
Target Name Vector Backbone Catalog Number
Oct-3/4 pMXs RTV-705
Sox2 pMXs RTV-706
c-Myc pMXs RTV-707
Klf4 pMXs RTV-708
NANOG pMXs RTV-711
Lin28 pMXs RTV-712
Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)
pMXs RTV-705-C
Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)
pMXs RTV-711-C
p53 shRNA pRetro RTV-400
Mouse iPS Vectors
Target Name Vector Backbone Catalog Number
Oct-3/4 pMXs RTV-701
Sox2 pMXs RTV-702
c-Myc pMXs RTV-703
Klf4 pMXs RTV-704
NANOG pMXs RTV-709
Lin28 pMXs RTV-710
Set of 4 vectors (Oct-3-4, Sox2, c-Myc, Klf4)
pMXs RTV-701-C
Set of 6 vectors (Oct-3-4, Sox2, c-Myc, Klf4, NANOG, Lin28)
pMXs RTV-709-C
p53 shRNA pRetro RTV-410
Human iPS Vectors
Product Name Size Catalog Number
Platinum-E Retroviral Packaging Cell Line, Ecotropic >3 x 106 cells RV-101
Platinum-A Retroviral Packaging Cell Line, Amphotropic >3 x 106 cells RV-102
Platinum-GP Retroviral Packaging Cell Line, Pantropic >3 x 106 cells RV-103
pCMV-VSV-G Packaging Vector (for use with Platinum-GP cells) 10 µg RV-110
Retroviral Packaging Cell Lines
*Kitamura, T. et al. (2003). Exp. Hematol. 31:1007-1014.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Induced Pluripotent Stem Cells
Polycistronic Vectors for iPS Cell Generation
STEM CELL RESEARCH
33
Product Name Size Catalog Number
pLentG-KOSM Polycistronic Lentiviral Vector (Mouse genes) 100 µL LTV-700
pRetroG-OKSM Polycistronic Retroviral Vector (Human genes) 100 µL RTV-700
Our Polycistronic Viral Vectors provide a convenient way to generate iPS cells. The defined stem cells factors Klf4, Oct-3/4, Sox2 and c-Myc are in-frame fused into a single open reading frame (ORF) by self-cleaving 2A peptides. The transcription factor ORF is followed by IRES-GFP as a reporter to verify viral transduction into your target cell. Efficiencies of iPS generation are typically higher compared to transduc-tion of four separate viruses each containing a single gene. Two vectors are available: pLentG-KOSM is a lentiviral vector containing
mouse sequences pRetroG-OKSM is a retroviral vector containing
human sequences
More Efficient: Up to 10-fold higher efficiency compared to multi-virus transduction, and 500-fold compared to non-viral methods
Reporter Convenience: GFP reporter gene helps to monitor viral transduction
Open Reading Frame of pLentG-KOSM Lentiviral Vector.
Characterization of iPS Cell Colonies Generated from MEFs Infected with Lentivirus Containing the KOSM Fusion. Top: Staining of pluripotency markers in induced cell colonies at 200x magnification. Bottom: AP staining at 100x magnification and mor-phology at 40x magnification in induced cell colonies.
Expression of Stem Cell Factors and GFP. Top: Transient ex-pression of KOSM fusion gene in 293T cells confirmed by Western blot. Bottom: GFP fluorescence in MEF cells 3 days after infection with lentivirus containing KOSM fusion.
For efficient packaging of your virus, please see our Lentiviral Packaging Systems on p. 58 and Retroviral Packaging Cell Lines on p. 64 and 67.
www.cellbiolabs.com [email protected]
Platinum Retroviral Expression Systems for Stem Cells
Product Name Size Catalog Number
Platinum ES/EC Retroviral Expression System, Ecotropic 1 kit VPK-303
Platinum ES/EC Retroviral Expression System, Amphotropic 1 kit VPK-304
1 kit VPK-305 Platinum ES/EC Retroviral Expression System, Pantropic
Platinum HSC Retroviral Expression System, Ecotropic 1 kit VPK-306
Platinum HSC Retroviral Expression System, Amphotropic 1 kit VPK-307
Platinum HSC Retroviral Expression System, Pantropic 1 kit VPK-308
34
Retroviral vectors are useful for delivering genes of interest into a host cell where integration into the genome is desired. However, traditional retroviral expression technologies usually result in low viral titers which make gene expression studies difficult.
Higher Viral Yields: Average titer 107 infec-tious units/mL with transient transfection
Versatile: 3 Packaging cell lines for use with nearly any target host species
Optimized for Stem Cell Studies: Specially designed expression systems for either ES/EC cells or hematopoietic stem cells
Amphotropic Ecotropic Pantropic
Human +++ N.S. +++
Mouse +++ +++ +++
Rat +++ +++ +++
Monkey +++ N.S. +++
Cat +++ N.S. +++
Dog +++ N.S. +++
Hamster + N.S. +++
Bird N.S. N.S. +++
Fish N.S. N.S. +++
Frog N.S. N.S. +++
Insect N.S. N.S. +++
Mollusk N.S. N.S. +++
*Virus must be packaged with a pantropic envelope protein such as VSVG. N.S. = Not Suitable
Catalog Number Packaging Cell Line Transfer Vector Envelope Vector Control Vector
VPK-303 Plat-E (Ecotropic) pMCs-Puro ——— pMCs-GFP
VPK-304 Plat-A (Amphotropic) pMCs-Puro ——— pMCs-GFP
VPK-305 Plat-GP (Pantropic) pMCs-Puro pCMV-VSV-G pMCs-GFP
VPK-306 Plat-E (Ecotropic) pMYs-Puro ——— pMYs-GFP
VPK-307 Plat-A (Amphotropic) pMYs-Puro ——— pMYs-GFP
VPK-308 Plat-GP (Pantropic) pMYs-Puro pCMV-VSV-G pMYs-GFP
Suitability of Platinum Retroviral Expression Systems by Host Species.
Components of the Platinum Retroviral Expression Systems for Stem Cells.
Our Platinum Retroviral Expression Systems incorpo-rate superior packaging cell lines and vector technolo-gies to produce high-titer virus with a single plasmid transfection. The Platinum Expression Systems in-clude one of our exclusive Platinum Packaging Cell Lines which already contain the gag and pol genes; the Ecotropic and Amphotropic cells also contain an envelope protein. Simply clone your gene of interest into the vector provided and transfect into the Plati-num cells. If you choose a Pantropic system, simply co-transfect with the VSV-G plasmid provided. The Platinum Expression Systems below are spe-cially designed for superior expression with either ES/EC cells or hematopoietic stem cells. For more infor-mation on our Platinum Expression Systems for a variety of cells, please see page 60.
STEM CELL RESEARCH Retroviral Expression Systems
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
MEF Feeder Cells
Product Name Size Catalog Number
MEF Feeder Cells 5 x 106 cells CBA-310
MEF Feeder Cells, Hygromycin-resistant 5 x 106 cells CBA-313
MEF Feeder Cells, Neomycin-resistant 5 x 106 cells CBA-311
MEF Feeder Cells, Puromycin-resistant 5 x 106 cells CBA-312
35
Our murine embryonic fibroblast (MEF) feeder cells are useful for the maintenance of human or mouse ES cells in their undifferentiated state. Cells must be mitotically inactivated prior to use.
Feeder Cells STEM CELL RESEARCH
SNL 76/7 Passage-Independent Feeder Cells for iPS Culture
The SNL 76/7 is an immortalized cell line derived from mouse fibroblast STO cells which have been transformed with murine LIF and neomycin resistance genes.
Stem Cell Feeders
Leukemia inhibitory factor (LIF) is useful for maintaining the undifferentiated state of mouse embryonic stem (mES) cells. However, LIF does not have the same effect on hu-man embryonic stem (hES) cells. Therefore, hES cells require the use of feeder cells for both derivation and maintenance. We offer a variety of feeder cells for stem cell culture. All feeder cells must be mitotically inactivated prior to use.
Superior Culture: Transformed with LIF gene for better maintenance of undifferentiated state
Versatile: Useful for culture of human and mouse iPS cells and as a feeder for ES cells
Passage-Independent: Immortalized cell line
Product Name Size Catalog Number
SNL Feeder Cells 3 x 106 cells CBA-316
Recent Product Citations 1. Osakada, F. et al (2009). In vitro differentiation of retinal cells
from human pluripotent stem cells by small-molecule induction. J. Cell Sci. 132:3169-3179.
2. Liu, Y. et al. (2009). Zeb1 represses Mitf and regulates pigment synthesis, cell proliferation, and epithelial morphology. Invest. Ophthalmol. Vis. Sci. 50:5080-5088.
3. Tsubooka, N. et al. (2009). Roles of Sall4 in the generation of pluripotent stem cells from blastocytes and fibroblasts. Genes Cells 14:683-694.
4. Lan, Z-J. et al. (2009). Extra-germ cell expression of mouse nuclear receptor subfamily 6, group A, member 1 (NR6A1). Biol. Reprod. 80:905-912.
JK1 Passage-Independent Feeder Cells
JK1 is an immortalized CD34+ stromal cell line that supports long-term proliferation of stem cells. It has been shown to maintain capacity for stem cell re-newal even after serial passaging for over one year. JK1 may be used for culture of a variety of cell types including pluripotent ES cells, germ-line derived stem cells such as SPCs and MASCs, and primordial germ cell-derived EG cells.
Product Name Size Catalog Number
JK1 Feeder Cells 1 x 106 cells CBA-315
JK1 Cells Support Maintenance of mES Cells. Immunofluores-cence staining of germ cell nu-clear antigen (GCNA). Antibody staining is green; nuclear coun-terstain is blue.
www.cellbiolabs.com [email protected]
36
CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay
Hematopoietic stem cells (HSCs), when cultured in a suitable semisolid matrix such as methylcellulose supplemented with cytokines & nutrients, proliferate to form discrete cell clusters or colonies. Such HSCs or hematopoietic progenitors are known as colony-forming cells (CFCs). In classic CFC assays, cells are cultured in a 35mm dish for 14-21 days so the colonies can reach a cer-tain size for manual counting, which can be tedious and subjective. The CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay provides a high-throughput method to quantify CFCs in just 7-10 days with no manual cell counting required. Cells are lysed, solu-bilized, and quantified using a fluorescent dye in-cluded in the kit. Alternatively, cells may be recov-ered for further culture and analysis.
Product Name Detection Size Catalog Number
CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay Fluorometric 96 Assays CBA-320
5 x 96 Assays CBA-320-5
Fast Results: 7-10 days vs. 2-3 weeks Plate Reader Convenience: Eliminates manual
counting Easier Reagent Handling: Methylcellulose media
can be handled using a pipet instead of a syringe
Assay Principle for the CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay.
HSC Colony Formation. Human bone marrow derived CD34+ Hematopoietic Progenitor Cells were seeded at 3000 cells/well and cultured for 7-10 days in the absence or presence of growth factors/cytokines. Colonies were quantified according to the assay protocol. A: After 7 days without cytokines. B: After 7 days in presence of cytokines. C: After 10 days in presence of cytokines (hemoglobin visible).
STEM CELL RESEARCH Colony Formation Assays
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
37
StemTAG™ 96-Well Stem Cell Colony Formation Assay
Product Name Detection Size Catalog Number
StemTAG™ 96-Well Stem Cell Colony Formation Assay Fluorometric 96 Assays CBA-325
5 x 96 Assays CBA-325-5
Our StemTAG™ 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantify ES cells in just 7-10 days with no manual cell count-ing required. After colonies are formed, stem cells may be ana-lyzed in 3 ways: 1. Lyse cells, then quantify using a fluorescent dye
included in the kit. 2. Lyse cells, then measure alkaline phosphate ac-
tivity using reagents provided. 3. Recover colonies for further culture and analysis. This assay may be of particular interest for the study of tumor stem cells.
Fast Results: 7-10 days vs. 2-3 weeks using con-ventional methods
Versatile: Quantify cells using fluorescent dye, measure alkaline phosphatase activity, or recover cells for further analysis
Plate Reader Convenience: No manual cell counting required
Anchorage-Independent Growth of Mouse ES-D3 Cells. Top: Phase Contrast. Bottom: Alkaline Phosphatase Staining.
StemTAG™ Stem Cell Colony Formation Assay Principle.
Colony Formation Assays STEM CELL RESEARCH
www.cellbiolabs.com [email protected]
StemTAG™ PCR Primer Set for Stem Cell Characterization
Product Name Size Catalog Number
StemTAG™ PCR Primer Set for Stem Cell Characterization 50 Reactions CBA-303
Total RNA and Protein from Murine ES-D3 Cell Line
Product Name Size Catalog Number
Total RNA—Murine Embryonic Stem Cell Line D3 50 µg CBA-304
Total Protein—Murine Embryonic Stem Cell Line D3 500 µg CBA-305
38
Pluripotent stem cells can differentiate into cells derived from all three embryonic germ layers: endoderm, mesoderm and ectoderm. Our StemTAG™ PCR Primer Set provides an efficient system for monitoring ES cell differentiation/undifferentiation. Seven primer sets are included: primers for two widely studied stem cell mark-ers (Oct-4 and NANOG), one marker for each embryonic germ layer (AFP/Endoderm, Flk-1/Mesoderm and NCAM/Ectoderm), and two controls (GAPDH and ß-Actin). Primers are suitable for either end-point or real-time (quantitative) PCR.
STEM CELL RESEARCH Alk Phos Assays, Primers, RNA/Protein
StemTAG™ Alkaline Phosphatase Kits
StemTAG™ Alkaline Phosphatase Staining Kit. Murine embry-onic stem cells (ES-D3) were maintained in an undifferentiated state with LIF. To induce differentiation, LIF was withdrawn over several days. Various differentiation events were observed: cells became flattened and enlarged with reduced proliferation. On day 5, cells were stained according to the assay protocol.
Product Name Detection Size Catalog Number
StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kit ICC & Colorimetric 2 x 100 Assays CBA-302
ICC & Fluorometric 2 x 100 Assays CBA-308
Colorimetric 100 Assays CBA-301
Fluorometric 100 Assays CBA-307
StemTAG™ Alkaline Phosphatase Staining Kit (Red) ICC 100 Assays CBA-300
StemTAG™ Alkaline Phosphatase Activity Assay Kit
StemTAG™ Alkaline Phosphatase Staining Kit (Purple) ICC 100 Assays CBA-306
The StemTAG™ Alkaline Phosphatase Staining and Activity Assay Kits monitor AP activity via both immu-nocytochemistry staining and a colorimetric 96-well plate-based activity assay. The staining and activity assay kits are also sold separately.
Fast Results: Staining and Activity Assay proto-cols each take less than 1 hour
Versatile: Useful for human ES, EG and EC cells, as well as mouse ES and EG cells
Recent Product Citations 1. Lee, J. et al. (2010). Ultraviolet A regulates adipogenic differen-
tiation of human adipose tissue-derived mesenchymal stem cells via up-regulation of kruppel-like factor 2. J. Biol. Chem. 285:32647-32656. (CBA-300)
2. Izadyar, F. et al. (2008). Generation of multipotent cell lines from a distinct population of male germ line stem cells. Reproduction 135:771-784. (CBA-300)
3. Kim, Y. et al. (2009). Cyclin-dependent kinase 2-associating protein 1 commits murine embryonic stem cell differentiation through retinoblastoma protein regulation. J. Biol. Chem. 284:23405-23414. (CBA-301)
4. Yue, Y. et al. (2008). A single intravenous injection of adeno-associated virus serotype-9 leads to whole body skeletal muscle transduction in dogs. Mol. Ther. 16(12):1944-1952. (CBA-301)
5. Ghosh, A. et al (2007). Efficient whole-body transduction with trans-splicing AAV vectors. Mol. Ther. 15(4):750-755. (CBA-301)
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Viral Vector Overview
Recombinant Viral Gene Delivery
Recombinant viral vectors provide a powerful means of delivering a gene into a target cell. There are many viral vectors available, and there are pros and cons to each. Use the fol-lowing table to select the best viral vector for your research.
Comparison of Viral Vectors for Gene Delivery Adeno-Associated
Virus (AAV) (p. 41-48)
Adenovirus (p. 49-56)
Lentivirus (HIV-1, FIV, SIV)
(p. 57-63)
Retrovirus (MMLV)
(p. 64-72)
Gene Expression Transient or Stable
Transient Transient or Stable
Stable
Will Infect Dividing Cells Yes Yes Yes Yes
Will Infect Non-Dividing Cells Yes Yes Yes No
Integrates into Target Cell Genome No* No Yes Yes
Relative Viral Titer XXX XXXX XXX XX
Relative Transduction Efficiency XXX XXXX XXX XX
Immune Response in Target Cells Very Low High Low Moderate
40
Typical Workflow for Viral Gene Delivery
Clone Gene of Interest
Package Virus
Measure Titer
Concentrate & Purify
Infect Target Cell
Viral Expression
Plasmid
Packaging Plasmids and Cells
Purification & Concentration
Kit
Viral Transduction
Reagents
Viral Quantitation
Kit
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
*Native AAV can integrate, but recombinant AAV rarely does.
Cell Biolabs offers kits and reagents for every step in your workflow.
VIRAL EXPRESSION AAV Expression
41
Adeno-Associated Virus Kits & Reagents
Adeno-associated virus (AAV) is less immunogenic than adenovirus or retrovirus. We offer a comprehensive line of AAV kits and reagents to ensure you get the best expression from your AAV expression studies:
AAV Helper Free Systems
AAV Helper Free Systems are available for a variety of formats and
serotypes: AAV Complete Expression Systems
contain all packaging plasmids plus an expression vector and a GFP control vector: p. 42-45
AAV Packaging Systems contain the pHelper plasmid and a serotype-specific Rep-Cap plasmid for use with your own expression construct: p. 45
If you have an AAV packaging system for one serotype and want to try another, choose one of 8 different AAV Rep-Cap Plasmids from native serotypes 1 through 6 plus AAV-DJ and AAV-DJ/8: p. 45
If you already have an AAV packaging system and need a cloning vector, choose one of 10 different AAV Expres-sion Vectors available individually: p. 46
Want to make a control virus? Choose one of our AAV Control Vectors: p. 46
Production of recombinant AAV requires certain genes from the adenovirus genome, which means that an adenovirus usually needs to be present. The AAV Helper Free System eliminates the need for a helper adenovirus. Most of the required adenoviral genes (E2A, E4 and VA RNA) are provided in a pHelper plasmid, while the required E1 gene is pro-vided by the 293 packaging cells.
Safer: pHelper plasmid eliminates the need for a helper virus
Flexible: Packaging vectors and expression vec-tors available separately or as one complete sys-tem, so you only order what you need
Expandable: All plasmids are provided individually, not in a mixture, so you can amplify in competent cells
Premade AAV Controls Purification Kits Quantitation / Titer Kit Transduction Kits
Helper Free Expression Systems Helper Free Packaging Systems Expression & Control Vectors Viral Packaging Cell Line
Gene Delivery using the AAV Helper Free System.
www.cellbiolabs.com [email protected]
VIRAL EXPRESSION AAV Expression
42 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Product Name Size Catalog Number
AAV-DJ Helper Free Expression System 1 kit VPK-410-DJ
AAV-DJ Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-DJ
AAV-DJ Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-DJ
AAV-DJ Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-DJ
AAV-DJ Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-DJ
AAV-DJ Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-DJ
AAV-DJ Helper Free Promoterless Expression System 1 kit VPK-411-DJ
AAV-DJ Helper Free shRNA Expression System (Puro) 1 kit VPK-412-DJ
AAV-DJ/Helper Free shRNA Expression System (GFP) 1 kit VPK-413-DJ
scAAV-DJ Helper Free Expression System 1 kit VPK-430-DJ
AAV Helper Free Complete Expression Systems
AAV Helper Free Complete Expression Systems con-tain everything you need to produce high-titer recom-binant adeno-associated virus:
pHelper Plasmid Rep-Cap Plasmid (serotype specific) GFP Control Vector Choice of 10 AAV Expression Vectors:
Gene Expression (CMV or no promoter) shRNA (U6 promoter) Self complementary (scAAV)
AAV Helper Free Expression Systems are available for the following serotypes: Native serotypes 1-6 AAV-DJ, engineered by DNA family shuffling to
form a hybrid capsid from 8 different native sero-types; provides significantly higher infectivity rates in vitro (see table below)
AAV-DJ/8, a mutant of AAV-DJ that exhibits in-creased uptake in brain and other tissues in vivo, similar to serotypes 8 and 9
Relative Infectivity Rates of AAV Serotypes. Normalized to AAV-2 = 100. ND = Not determined.
Cell Line Cell or Tissue Source AAV-1 AAV-2 AAV-3 AAV-4 AAV-5 AAV-6 AAV-8 AAV-9 AAV-DJ
AAV-DJ/8
Huh-7 Hu Liver 13 100 2.5 0.0 0.1 10 0.7 0.0 500 0.2
HEK293 Hu Kidney 25 100 2.5 0.1 0.1 5 0.7 0.1 500 0.3
HeLa Hu Cervix 3 100 2.0 0.1 3.7 1.0 0.2 0.1 667 0.2
HepG2 Hu Liver 3 100 16.7 0.3 1.7 5 0.3 ND 1250 0.5
Hep1A Ms Liver 20 100 0.2 1.0 0.1 1.0 0.2 0.0 400 0.1
911 Hu Retina 17 100 11.1 0.2 0.1 17 0.1 ND 500 0.0
CHO Hm Ovary 100 100 14.3 1.4 333 50 10.0 1.0 25000 5.0
COS Si Kidney 33 100 33 3.3 5.0 14 2.0 0.5 500 0.3
MeWo Hu Skin 10 100 20 0.3 6.7 10 1.0 0.2 2857 1.0
NIH3T3 Ms Fibroblasts 10 100 2.9 2.9 0.3 10 0.3 ND 500 0.1
A549 Hu Lung 14 100 20 ND 0.5 10 0.5 0.1 1000 0.1
HT1180 Hu Fibroblasts 20 100 10.0 0.1 0.3 33 0.5 0.1 333 0.2
Monocytes Hu Primary Monocytes 1111 100 ND ND 125 1429 ND ND 100 ND
Immature DC Hu Monocyte-derived DC 2500 100 ND ND 222 2857 ND ND 200 ND
Mature DC Hu Monocyte-derived DC 2222 100 ND ND 333 3333 ND ND 100 ND
AAV-DJ Helper Free Complete Expression Systems
AAV-DJ/8 Helper Free Complete Expression Systems
VIRAL EXPRESSION AAV Expression
43 www.cellbiolabs.com [email protected]
AAV-1 Helper Free Complete Expression Systems
Product Name Size Catalog Number
AAV-DJ/8 Helper Free Expression System 1 kit VPK-410-DJ-8
AAV-DJ/8 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-DJ-8
AAV-DJ/8 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-DJ-8
AAV-DJ/8 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-DJ-8
AAV-DJ/8 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-DJ-8
AAV-DJ/8 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-DJ-8
AAV-DJ/8 Helper Free Promoterless Expression System 1 kit VPK-411-DJ-8
AAV-DJ/8 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-DJ-8
AAV-DJ/8 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-DJ-8
scAAV-DJ/8 Helper Free Expression System 1 kit VPK-430-DJ-8
Product Name Size Catalog Number
AAV-1 Helper Free Expression System 1 kit VPK-410-SER1
AAV-1 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER1
AAV-1 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER1
AAV-1 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER1
AAV-1 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER1
AAV-1 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER1
AAV-1 Helper Free Promoterless Expression System 1 kit VPK-411-SER1
AAV-1 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER1
AAV-1 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER1
scAAV-1 Helper Free Expression System 1 kit VPK-430-SER1
AAV-2 Helper Free Complete Expression Systems
Product Name Size Catalog Number
AAV-2 Helper Free Expression System 1 kit VPK-410-SER2
AAV-2 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER2
AAV-2 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER2
AAV-2 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER2
AAV-2 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER2
AAV-2 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER2
AAV-2 Helper Free Promoterless Expression System 1 kit VPK-411-SER2
AAV-2 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER2
AAV-2 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER2
scAAV-2 Helper Free Expression System 1 kit VPK-430-SER2
VIRAL EXPRESSION AAV Expression
44 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
AAV-3 Helper Free Complete Expression Systems
AAV-4 Helper Free Complete Expression Systems
Product Name Size Catalog Number
AAV-3 Helper Free Expression System 1 kit VPK-410-SER3
AAV-3 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER3
AAV-3 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER3
AAV-3 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER3
AAV-3 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER3
AAV-3 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER3
AAV-3 Helper Free Promoterless Expression System 1 kit VPK-411-SER3
AAV-3 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER3
AAV-3 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER3
scAAV-3 Helper Free Expression System 1 kit VPK-430-SER3
Product Name Size Catalog Number
AAV-4 Helper Free Expression System 1 kit VPK-410-SER4
AAV-4 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER4
AAV-4 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER4
AAV-4 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER4
AAV-4 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER4
AAV-4 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER4
AAV-4 Helper Free Promoterless Expression System 1 kit VPK-411-SER4
AAV-4 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER4
AAV-4 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER4
scAAV-4 Helper Free Expression System 1 kit VPK-430-SER4
AAV-5 Helper Free Complete Expression Systems
Product Name Size Catalog Number
AAV-5 Helper Free Expression System 1 kit VPK-410-SER5
AAV-5 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER5
AAV-5 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER5
AAV-5 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER5
AAV-5 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER5
AAV-5 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER5
AAV-5 Helper Free Promoterless Expression System 1 kit VPK-411-SER5
AAV-5 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER5
AAV-5 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER5
scAAV-5 Helper Free Expression System 1 kit VPK-430-SER5
VIRAL EXPRESSION AAV Expression
45 www.cellbiolabs.com [email protected]
AAV-6 Helper Free Complete Expression Systems
Product Name Size Catalog Number
AAV-6 Helper Free Expression System 1 kit VPK-410-SER6
AAV-6 Helper Free Bicistronic Expression System (Puro) 1 kit VPK-415-SER6
AAV-6 Helper Free Bicistronic Expression System (Neo) 1 kit VPK-416-SER6
AAV-6 Helper Free Bicistronic Expression System (Hygro) 1 kit VPK-417-SER6
AAV-6 Helper Free Bicistronic Expression System (GFP) 1 kit VPK-418-SER6
AAV-6 Helper Free Bicistronic Expression System (Blasticidin) 1 kit VPK-419-SER6
AAV-6 Helper Free Promoterless Expression System 1 kit VPK-411-SER6
AAV-6 Helper Free shRNA Expression System (Puro) 1 kit VPK-412-SER6
AAV-6 Helper Free shRNA Expression System (GFP) 1 kit VPK-413-SER6
scAAV-6 Helper Free Expression System 1 kit VPK-430-SER6
AAV Helper Free Packaging Systems
AAV Helper Free Packaging Systems contain everything found in the Complete Expression Systems, with the exception of the AAV expression vector. This is an ideal choice if you already have an AAV construct containing your gene of interest. All plasmids are provided individually, not as a packaging mixture.
Product Name Size Catalog Number
AAV-2 Helper Free Packaging System 1 kit VPK-402
AAV-1 Helper Free Packaging System 1 kit VPK-401
AAV-3 Helper Free Packaging System 1 kit VPK-403
AAV-4 Helper Free Packaging System 1 kit VPK-404
AAV-DJ Helper Free Packaging System 1 kit VPK-400-DJ
AAV-DJ/8 Helper Free Packaging System 1 kit VPK-400-DJ-8
AAV-5 Helper Free Packaging System 1 kit VPK-405
AAV-6 Helper Free Packaging System 1 kit VPK-406
AAV Rep-Cap Plasmids (Serotype-Specific)
AAV Rep-Cap plasmids allow you to make recombinant AAV of a specific serotype. These plasmids are ideal if you already have an AAV packaging system for a different serotype. Just substitute one of these plasmids into your AAV Helper Free Packaging System or Expression System.
Product Name Catalog Number
pAAV-DJ Vector VPK-420-DJ
pAAV-DJ/8 Vector VPK-420-DJ-8
pAAV-RC1 Vector VPK-421
pAAV-RC2 Vector VPK-422
Product Name Catalog Number
pAAV-DJ Vector VPK-420-DJ
pAAV-DJ/8 Vector VPK-420-DJ-8
pAAV-RC1 Vector VPK-421
pAAV-RC2 Vector VPK-422
VIRAL EXPRESSION AAV Expression
46
AAV Expression Vectors
Product Name Size Catalog Number
pAAV-MCS Expression Vector 10 µg VPK-410
pAAV-IRES-Puro Expression Vector 10 µg VPK-415
pAAV-IRES-Neo Expression Vector 10 µg VPK-416
pAAV-IRES-Hygro Expression Vector 10 µg VPK-417
pAAV-IRES-GFP Expression Vector 10 µg VPK-418
pAAV-IRES-Bsd Expression Vector 10 µg VPK-419
pAAV-MCS Promoterless Expression Vector 10 µg VPK-411
pAAV-U6-Puro Expression Vector 10 µg VPK-412
pAAV-U6-GFP Expression Vector 10 µg VPK-413
Cloning Capacity
3 kb
1.8 kb
1.6 kb
1.4 kb
1.7 kb
2 kb
3.9 kb
2.2 kb
2.1 kb
pscAAV-MCS Expression Vector 1.5 kb 10 µg VPK-430
AAV Control Plasmids
Product Name Size Catalog Number
pAAV-GFP Control Vector 10 µg AAV-400
pAAV-Cre Control Vector 10 µg AAV-401
pAAV-LacZ Control Vector 10 µg AAV-402
pscAAV-GFP Control Vector 10 µg AAV-410
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
AAV Premade Control Viruses
Product Name Size Catalog Number
AAV1-GFP Control Virus 50 µL AAV-301
AAV2 Null Control Virus 50 µL AAV-300
AAV2-Cre Control Virus 50 µL AAV-310
AAV2-GFP Control Virus 50 µL AAV-302
AAV2-Luc Control Virus 50 µL AAV-320
AAV3-GFP Control Virus 50 µL AAV-303
AAV5-GFP Control Virus 50 µL AAV-305
AAV6-GFP Control Virus 50 µL AAV-306
All AAV premade viruses are provided at a concentration of 1 x 1012 GC/mL.
Each of our AAV Expression Vectors may be used with any of our AAV Helper Free Systems, regardless of AAV serotype. Choose one of these vectors when you already have an AAV Packaging System but may want to use a different promoter or selection marker.
Choose one of our AAV control vectors when you already have an AAV Packaging System and want to make a transduction control virus.
Recent Product Citation Sen, Y. et al. (2014). TDP-43 causes differential pathology in neu-ronal versus glial cells in the mouse brain. Human Mol. Genet. 10.1093/hmg/ddt662. (VPK-410)
VIRAL EXPRESSION AAV Expression
47
ViraBind™ AAV Purification Kits
Purification of AAV via ultracentrifugation can be tedious and time-consuming, and may result in low yields. ViraBind™ AAV Purification Kits use a one-step proprietary matrix followed by further purification and concen-tration using a centrifugal concentrator. The result is a higher AAV yield with high purity in a fraction of the time. Kits are suitable for AAV-2 or AAV-DJ; they will not work with other AAV serotypes.
High Purity: No contamination bands as seen on SDS gel
Fast Results: Obtain purified virus in about 3 hours
High Yields: Recovery rate >60%
Electrophoretic Profile of Purified AAV2-GFP.
Product Name Capacity/Prep Size Catalog Number
ViraBind™ AAV Purification Kit Two 10-cm dishes 10 Preps VPK-140
ViraBind™ AAV Purification Mega Kit 2 Preps VPK-141
10 Preps VPK-141-5 Ten 15-cm dishes
Purification Procedure for the ViraBind™ AAV Purification Kit.
293AAV Cell Line
Product Name Size Catalog Number
1 x 106 cells AAV-100 293AAV Cell Line
Our 293AAV cell line is selected from the parental 293 cell line for larger surface area, flattened morphology, and firmer attachment to culture plates, resulting in production of higher yields of AAV.
www.cellbiolabs.com [email protected]
Recent Product Citation Uchida, S. et al. (2010). Early life stress enhances behavioral vulnerability to stress through the activation of REST4-mediated gene transcription in the medial prefrontal cortex of rodents. J. Neurosci. 30:15007-15018. (VPK-140)
VIRAL EXPRESSION AAV Expression
QuickTiter™ AAV Quantitation Kit
Product Name Capacity/Prep Size Catalog Number
QuickTiter™ AAV Quantitaiton Kit Fluorometric 20 Assays VPK-145
Fast Results: Obtain purified virus in less than 2 hours
High Sensitivity: Limit of detection 1 x 109 GC/mL from unpurified supernatant or 5 x 1010 GC/mL from purified AAV
Traditional AAV Quantitation by dot blot can be tedi-ous, time consuming, and suffer from high inter-assay variability. Our QuickTiter™ AAV Quantitation Kit uses a proprietary technology to quantify AAV nucleic acid content of unpurified AAV-2 or AAV-DJ, or from purified AAV of any serotype.
0
25
50
75
100
125
150
175
200
0 25 50 75 100 125 150
AAV DNA STD (ng)
RF
U
0
5
10
15
20
0 2 4 6 8 10
AAV DNA STD (ng)
RF
U
AAV-2 DNA Standard Curve. The QuickTiter™ AAV-2 DNA Standard was diluted as described in the assay protocol. Fluorescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485/538 nm filter set and 530 nm cutoff.
ViraDuctin™ AAV Transduction Reagent
Product Name Size* Catalog Number
10 Transductions AAV-200
50 Transductions AAV-201 ViraDuctin™ AAV Transduction Reagent
48
*Number of transductions performed in 35mm culture dishes. May be modified for use in culture plates or larger dishes. See product insert.
Our ViraDuctin™ AAV Transduction Reagent can significantly increase the transduction efficiency of AAV vectors in both dividing and non-dividing cells. Increases are greatest in non-dividing cells, but even cells in S-phase show a noticeable increase in trans-duction efficiencies.
Higher Efficiencies: Significantly increase rate of infection of host cells
Low Toxicity: No noticeable effect on cell viability Universal: Suitable for use with both dividing and
non-dividing cells
Successful gene expression studies using AAV depend on high transduction efficiencies into host cells. Infection rates appear to be highest in S-phase cells, which can account for a very small fraction of a cell population.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Adenoviral Expression
49
Adenoviral Expression Kits & Reagents
Recombinant adenoviruses are excellent tools for introducing genetic material into host cells, since they can infect a variety of mammalian cell types with high efficiency. They re-main epichromosal upon infection, so they are only suitable for transient gene expression. We offer a complete workflow solution to your adenoviral expression studies:
Purification Kits Quantitation / Titer Kits Transduction Reagents
Viral Expression Systems Viral Packaging Cell Line Premade Recombinant Adenoviruses
RAPAd® Adenoviral Expression Systems
Adenovirus Production using the RAPAd® Adenoviral Expression System.
Compared to other adenoviral expression systems, RAPAd® Adenoviral Expression Systems produce recombinant adenovirus in a much shorter time (about 2-3 weeks) with a substantial reduction in wild-type adenovirus. The RAPAd systems use a backbone vector from which the 5’ ITR, packaging signal and E1 sequences have been removed. Addi-tionally, serial amplification of the recombinant ade-novirus does not increase the level of replication-competent adenovirus.
Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods
Faster Production: Virus generated in 2-3 weeks compared to a few months with traditional methods
7 Complete Systems: Choose CMV or RSV for gene expression, EF-1 for miRNA expression, U6 for shRNA, or clone your own promoter along with your gene of interest using our Universal system
Product Name Promoter Size Catalog Number
RAPAd® Universal Adenoviral Expression System None 1 Kit VPK-250
RAPAd® RSV Adenoviral Expression System RSV 1 Kit VPK-251
RAPAd® CMV Adenoviral Expression System CMV 1 Kit VPK-252
RAPAd® miRNA Adenoviral Expression System EF-1 1 Kit VPK-253
RAPAd® Bicistronic Adenoviral Expression System (GFP) CMV 1 Kit VPK-254
RAPAd® shRNA Adenoviral Expression System (Puro) U6 1 Kit VPK-255
RAPAd® shRNA Adenoviral Expression System (GFP) U6 1 Kit VPK-256
www.cellbiolabs.com [email protected]
Recent Product Citations 1. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond
is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (VPK-252)
2. Li, P. et al. (2013). MicroRNA-638 is highly expressed in hu-man vascular smooth muscle cells and inhibits PDGF-BB-induced cell proliferation and migration through targeting or-phan nuclear receptor NOR1. Cardiovasc. Res. 10.1093/cvr/cvt082. (VPK-253)
50
VIRAL EXPRESSION Adenoviral Expression
Don’t have time to make your own adenovirus? Are you studying the expression of multiple genes? Rely on our premade recombinant adenoviruses that al-ready contain a gene of interest. All of Cell Biolabs’ premade recombinant adenoviruses are provided as 50 µl aliquots at a concentration of 1 x 1011 viral parti-cles/mL in TBS with 10% glycerol.
Target Name Catalog Number
Null Control (No gene) ADV-001
-Galactosidase ADV-002
Cre ADV-005
Firefly Luciferase ADV-008
GFP ADV-004
SEAP (Secretory Alkaline Phosphatase) ADV-003
Controls and Reporter Genes
Recent Product Citations 1. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis
closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-001)
2. Zhang, Z. et al (2013). MEK inhibition leads to lysosome-mediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV-002)
3. Schramm, C. et al (2012). The PTPN11 loss-of-function muta-tion Q510E-Shp2 causes hypertrophic cardiomyopathy by dys-regulating mTOR signaling. Am. J. Physiol. Heart Circ. Physiol. 302:H231-H243. (ADV-002)
4. Salvati, E. et al. (2014). Evidence for G-quadruplex in the pro-moter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-004)
5. Lu, D. et al. (2012). Peroxisome proliferator-activated receptor-coactivator-1alpha enhances engraftment and angiogenesis of mesenchymal stem cells in diabetic hindlimb ischemia. Diabe-tes 61:1153-1159. (ADV-004)
6. Kato, H. et al. (2011). Wnt/ß-Catenin pathway in podocytes integrates cell adhesion, differentiation and survival. J. Biol. Chem. 286:26003-26015. (ADV-005)
Premade Recombinant Adenoviruses
Cancer/Tumor Antigens
Target Name Catalog Number
Carbonic Anhydrase 9 (CA9) ADV-602
Carcinoembryonic Antigen (CEA) ADV-604
NY-ESO-1 ADV-601
Blood Vessel Formation After 3 Days. Purified Ad-Null or Ad-VEGF viruses were applied to a 10-day old CAM (chick chorioal-lanoic membrane). Results were visualized by stereomicroscope.
Angiogenesis
Target Name Catalog Number
VEGF ADV-101
HIF-1 ADV-100
Recent Product Citations 1. Kelber, J.A. et al. (2012). KRas induces a Src/PEAK1/ErbB2
kinase amplification loop that drives metastatic growth and therapy resistance in pancreatic cancer. Cancer Res. 72:2554-2564. (ADV-101)
2. Qiu, X. et al. (2012). Combined strategy of mesenchymal stem cell injection with vascular endothelial growth factor gene ther-apy for the treatment of diabetes-associated erectile dysfunc-tion. J. Androl. 33:37-44. (ADV-101)
3. Stoletov, K. et al. (2010). Visualizing extravasation dynamics of metastatic tumor cells. J. Cell Sci. 123:2332-2341. (ADV-101)
4. Serban, D. et al. (2008). H-ras regulates angiogenesis and vascular permeability by activation of distinct downstream ef-fectors. Circ. Res. 102(11):1350-1358. (ADV-101)
293AD Cell Line for Adenoviral Packaging and Amplification
The 293AD cell line is derived from the parental 293 cell line, but has been specifically selected for adeno-virus applications and offers advantages over conven-tional 293 cells: flattened morphology, firm attach-ment to culture plates, and a larger surface area for superior transfection and greater viral yields.
Recent Product Citations 1. Peng, D. et al. (2011). Glutathione peroxidase 7 protects
against oxidative DNA damage in oesophageal cells. Gut 61:1250-1260. (AD-100)
2. Kothari, H. et al. (2010). Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption. Blood 115:4273-4283. (AD-100)
Product Name Size Catalog Number
293AD Cell Line 1 x 106 Cells AD-100
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Adenoviral Expression
51
Premade Recombinant Adenoviruses, continued
Actin Cytoskeleton Staining. Cos-7 cells were infected with puri-fied Ad-Ras V12 (ADV-146) at 50 MOI (multiplicity of infection). Membrane ruffling was visualized by staining the actin cytoskeleton with Rhodamine-coupled Phalloidin.
Cytoskeleton Regulation / Small GTPase
Target Name Catalog Number
Cdc42 ADV-152
Cdc42 L61 (Constitutively Active) ADV-154
Cdc42 N17 (Dominant Negative) ADV-153
PAK1 ADV-202
PAK1 (H83L, H86L) ADV-203
PAK1 (H83L, H86L, K299R) ADV-205
PAK1 (K299R) ADV-207
PAK1 (L107E, T423E) ADV-206
PAK1 (T423E) ADV-204
PAK1 (Kinase Domain) ADV-209
PAK1 (Regulatory Domain) ADV-208
Rac1 ADV-149
Rac1 L61 (Constitutively Active) ADV-151
Rac1 N17 (Dominant Negative) ADV-150
Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue
growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847. (ADV-145, ADV-150, ADV-153, ADV-156)
2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150)
3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150)
4. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154)
5. Salvati, E. et al. (2014). Evidence for G-quadruplex in the pro-moter of vegfr-2 and its targeting to inhibit tumor angiogenesis. Nucleic Acids Res. 42:2945-2957. (ADV-151, ADV-157)
6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42-dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153)
7. Neal M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)
8. Nie, J. et al. (2013). SAD-A kinase controls islet ß-cell size and function as a mediator of mTORC1 signaling. PNAS 110:13857-13862. (ADV-204, ADV-207)
9. Nie, J. et al. (2013). Synapses of amphids defective (SAD-A) kinase promotes glucose-stimulated insulin secretion through activation of p21-activated kinase (PAK1) in pancreatic ß-cells. J. Biol. Chem. 287:26435-26444. (ADV-204, ADV-207)
Cell Cycle & Transcription Regulation
Target Name Catalog Number
p53 ADV-501
p53 (Temperature Sensitive Mutant) ADV-502
p68 RNA Helicase ADV-505
Myogenin ADV-509
Target Name Catalog Number
DCC ADV-504
MyoD ADV-508
Target Name Catalog Number
Ras N17 (Dominant Negative) ADV-145
Ras V12 (Constitutively Active) ADV-146
Ras V12C40 ADV-148
Ras V12S35 ADV-147
Rho L63 (Constitutively Active) ADV-157
Rho N19 (Dominant Negative) ADV-156
SDF-1 ADV-210
www.cellbiolabs.com [email protected]
Recent Product Citation Nguyen, N. et al. (2014). Mitsugumin 53 (MG53) ligase ubiquiti-nates focal adhesion kinase during skeletal myogenesis. J. Biol. Chem. 289:3209-3216. (ADV-508)
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VIRAL EXPRESSION Adenoviral Expression
MAP Kinase Signaling
Premade Recombinant Adenoviruses, continued
Target Name Catalog Number
Cdc42 ADV-152
Cdc42 L61 (Constitutively Active) ADV-154
Cdc42 N17 (Dominant Negative) ADV-153
ERK2 ADV-112
ERK2 (Dominant Negative) ADV-113
ERK5 (BMK1) ADV-116
ERK5 (Dominant Negative) ADV-117
Interferon- ADV-103
Interleukin-2 ADV-102
JNK1 ADV-114
JNK1 (Dominant Negative) ADV-115
MAPKAPK2 ADV-137
MAPKAPK2 (Dominant Negative) ADV-138
MAPKAPK2 (Constitutively Active) ADV-139
MEK1 (Dominant Negative) ADV-118
MEK1 (Constitutively Active) ADV-119
MEK5 ADV-129
MEK5 (Dominant Negative) ADV-130
MEK5 (Constitutively Active) ADV-131
MEKK1 ADV-135
MEKK1 (Dominant Negative) ADV-136
MEKK3 ADV-162
Immunoblot of Various Cell Signaling Targets. HUVEC cells were in-fected with purified Ad-Null (ADV-001), Ad-Ras V12 (ADV-146), Ad-Ras V12S35 (ADV-147), Ad-Ras V12C40 (ADV-148), Ad-MEK1 (ADV-119), and Ad-Rac1 L61 (ADV-151) at 10 MOI (multiplicity of infection). Cell lysates were ana-lyzed for gene expres-sion and ERK activation.
Target Name Catalog Number
Rac1 ADV-149
Rac1 L61 (Constitutively Active) ADV-151
Rac1 N17 (Dominant Negative) ADV-150
Raf1 ADV-132
Raf1 (Dominant Negative) ADV-133
Raf1 (Constitutively Active) ADV-134
Ras N17 (Dominant Negative) ADV-145
Ras V12 (Constitutively Active) ADV-146
Rho L63 (Constitutively Active) ADV-157
Rho N19 (Dominant Negative) ADV-156
SOK ADV-142
SOK (Dominant Negative) ADV-143
SOK (Constitutively Active) ADV-144
Tac-Rac1 (Membrane Targeting) ADV-164
MKK6 (Dominant Negative) ADV-124
MKK6 (Constitutively Active) ADV-125
MKK7 ADV-126
MKK7 (Dominant Negative) ADV-127
MKK7 (Constitutively Active) ADV-128
myr-Rac1 ADV-163
p38 ADV-104
p38 (Dominant Negative) ADV-105
p38 ADV-106
p38 (Dominant Negative) ADV-107
p38 ADV-108
p38 (Dominant Negative) ADV-109
p38 (Dominant Negative) ADV-111
PRAK (Dominant Negative) ADV-141
MKK3 (Dominant Negative) ADV-121
MKK3 (Constitutively Active) ADV-122
MKK4 (Dominant Negative) ADV-160
MKK4 (Constitutively Active) ADV-161
MKK6 ADV-123
MKK3 ADV-120
See following page for recent citations using these adenoviruses
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
53
VIRAL EXPRESSION Adenoviral Expression
Premade Recombinant Adenoviruses, continued
Tyrosine Kinases and PKCs
Target Name Catalog Number
CSK ADV-405
CSK (Dominant Negative) ADV-406
Fyn ADV-403
Fyn (Dominant Negative) ADV-404
PKC- (Dominant Negative) ADV-410
PKC- (Dominant Negative) ADV-411
PKC- (Dominant Negative) ADV-412
shAkt1 ADV-417
shAkt2 ADV-418
Src ADV-401
Recent Product Citations 1. Harbrecht, B.G. et al. (2012). Insulin inhibits hepatocyte iNOS
expression induced by cytokines by an Akt-dependent mecha-nism. Am. J. Physiol. Gastrointest. Liver Physiol 302:G116-G122. (ADV-105)
2. Jones, S.W. et al. (2009). Mitogen-activated protein kinase-activated protein kinase (MK2) modulates key biological path-ways associated with OA disease pathology. Osteoarthritis and Cartilage 17:124-131. (ADV-105)
3. Kim, J.M. et al. (2008). Inhibition of apoptosis in Bacteroids fragilis enterotoxin-stimulated intestinal epithelial cells through the induction of c-IAP-2. Eur. J. Immunol. 38(8):2190-2199. (ADV-105)
4. Lee, J.Y. et al. (2007). Effects of transcription factor activator protein-1 on interleukin-8 expression and enteritis in response to Clostridium difficile toxin A. J. Mol. Med. 85:1393-1404. (ADV-105, ADV-115)
5. Monick, M. et al. (2008). Constitutive ERK MAPK activity regu-lates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496. (ADV-112, ADV-113, ADV-118, ADV-119)
6. Samuel, I. et al. (2008). Enteral exclusion increases MAP kinase activation and cytokine production in a model of gall-stone pancreatitis. Pancreatology 8(1):6-14. (ADV-113)
7. Wang, X. et al. (2007). Human immunodeficiency virus prote-ase inhibitor ritonavir inhibits cholesterol efflux from human macrophage-derived foam cells. Am. J. of Pathology 171:304-314. (ADV-113)
8. Jiang, S. et al. (2011). Role of inhibitory kB kinase and c-Jun NH2-terminal kinase in the development of hepatic insulin re-sistance in critical illness diabetes. Am. J. Physiol. Gastrointest. Liver Physiol 301:G454-G463. (ADV-115)
9. Zhang, Z. et al. (2013). MEK inhibition leads to lysosome-mediated Na+/I– symporter protein degradation in human breast cancer cells. Endocr. Relat. Cancer 20:241-250. (ADV-118)
10. Matsushita, T. et al. (2009). FGFR3 promotes synchondosis closure and fusion of ossification centers through the MAPK pathway. Hum. Mol. Genet. 18:227-240. (ADV-119)
11. Yoon, C-H. et al. (2009). Activation of p38 mitogen-activated protein kinase is required for death receptor-independent cas-pase-8 activation and cell death in response to sphingosine. Mol. Cancer Res. 7(3):361-370. (ADV-119)
12. Tan, S.H. et al. (2009). Regulation of cell proliferation and mi-gration by TAK1 via transcriptional control of von Hippel-Lindau tumor suppressor. J. Biol. Chem. 284:18047-18058. (ADV-128)
13. Wu, Y. et al. (2012). ERK5 regulates glucose-induced in-creased fibronectin production in the endothelial cells and in the retina in diabetes. Invest. Ophthalmol. Vis. Sci. 53:8405-8413. (ADV-130)
14. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires vß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Cell Biol. 23:1104-1114. (ADV-150)
15. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150)
16. Neal, M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)
17. Taniguchi, C. et al. (2007). The p85a regulatory subunit of phosphoinositide 3-kinase potentiates c-Jun N-terminal kinase-mediated insulin resistance. Mol. Cell Biol. 27:2830-2840. (ADV-161)
MAP Kinase Signaling, continued NFB Signaling
Target Name Catalog Number
IB- ADV-301
IB- S32A (Dominant Negative) ADV-302
IKK- ADV-305
IKK- (Dominant Negative) ADV-303
NOD2 ADV-308
Rel B ADV-304
Recent Product Citations 1. Johnston, R.K. et al. (2009). ß3-integrin mediated ubiquitination
activates survival signaling during myocardial hypertrophy. FASEB J. 23(8):2759-2771. (ADV-302)
2. Martin, A.P. et al. (2008). Lapatinib resistance in HCT116 cells is mediated by elevated MCL-1 expression and decreased BAK activation and not by ERBB receptor kinase mutation. Mol. Pharmacol. 74:807-822. (ADV-302)
3. Richardson, W.M. et al. (2010). Nucleotide-binding oligomeri-zation domain-2 inhibits toll-like receptor-4 signaling in the intestinal epithelium. Gastroenterology 139(3):904-917. (ADV-308, ADV-309)
Recent Product Citations 1. Koh, W. et al. (2009). Formation of endothelial lumens requires a
PKC-, Src-, Pak-, and Raf-kinase dependent signaling cascade downstream of Cdc42 activation. J. Cell Sci. 122:1812-1822. (ADV-401, ADV-405, ADV-406)
2. Lecuona, E. et al. (2009). Ubiquitination participates in the ly-sosomal degradation of the Na,K-ATPase in steady state condi-tions. Am. J. Respir. Cell Mol. Biol. 41(6):617-679. (ADV-412)
3. Vadasz, I. et al. (2008). AMP-activated protein kinase regulates CO2-induced alveolar epithelial dysfunction in rats and human cells by promoting Na,K-ATPase endocytosis. J. Clin. Invest. 118(2):752-762. (ADV-412)
4. Briva, A. et al. (2007). High CO2 levels impair alveolar epithelial function independent of pH. PLoS ONE 2(11):e1238. (ADV-412)
www.cellbiolabs.com [email protected]
VIRAL EXPRESSION Adenoviral Expression
54
ViraBind™ Adenovirus Purification Kits
Purification of Recombinant Ad--Gal. Ad-β-Gal was purified according to the assay protocol. Each purification fraction was used to infect A549 cells in a 12-well plate. After 48 hr, cells were scored using our β-Galactosidase Staining Kit (p. 92).
Product Name Capacity/Prep Size Catalog Number
ViraBind™ Adenovirus Miniprep Kit 1 x 1011 VP 10 Preps VPK-099
ViraBind™ Adenovirus Purification Kit 2.5 x 1012 VP 10 Preps VPK-100
Purification of viruses via cesium chloride (CsCl) ul-tracentrifugation procedures can be tedious and time-consuming. ViraBind™ Adenovirus Purification Kits use an efficient system for quick adenoviral purifica-tion with high recovery. No ultracentrifugation is re-quired. Kits use either a spin column or syringe filter for high purity adenovirus (see selection guide).
High Viral Yield: >90% recovery High Quality: Provides quality of CsCl procedures,
but in much less time Faster Results: 30 minutes (1-2 hrs for Mega kit) User-Friendly Protocol: No gradient preparation
or ultracentrifugation steps
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Recent Product Citations 1. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a
key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-099)
2. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal forma-tion. Cardiovasc. Res. 95:517-526. (VPK-099)
3. Kirui, J.K. et al. (2010). Gßgamma signaling promotes breast cancer cell migration and invasion. J. Pharmacol. Exp. Ther. 333:393-403. (VPK-099)
4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-100)
5. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-100)
6. Chen, F. et al. (2011). Dynamic regulation of PDX-1 and FoxO1 expression by FoxA2 in dexamethasone-induced pan-creatic ß-cells dysfunction. Endocrinology 152:1779-1788. (VPK-100)
7. Prasad, S.S. et al. (2011). Enzymatic activities of the human AGPAT isoform 3 and isoform 5: localization of AGPAT5 to mitochondria. J. Lipid Res. 52:451-462. (VPK-100)
8. Agarwal, A.K. et al. (2010). Enyzmatic activity of the human 1-acylglycerol-3-phosphate-O-acyltransferase isoform 11: upregulated in breast and cervical cancers. J. Lipid Res. 51:2143-2152. (VPK-100)
9. Triulzi, C. et al. (2010). Antibody-dependent natural killer cell-mediated cytotoxicity engendered by a kinase-inactive HER2 adenovirus-based vaccination mediates resistance to breast tumors. Cancer Res. 70:7431-7441. (VPK-100)
10.Sen, P. et al. (2010). Zinc modulates the interaction of protein C and activated protein C with endothelial protein C receptor. J. Biol. Chem. 285:20410-20420. (VPK-100)
11.Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pan-creatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (VPK-100)
12.Lam, Y.W. et al. (2010). Proteomics analysis of the nucleolus in adenovirus-infected cells. Mol. Cell. Proteomics 9:117-130. (VPK-100)
Selection Guide for ViraBind™ Adenovirus Purification Kits
ViraBind™ Adenovirus Miniprep Kit
ViraBind™ Adenovirus Purification Kit
Purification Method Spin column Syringe filter
Purification Time 30 minutes 30 minutes
Capacity/Prep (Viral Particles) 1 x 1011 VP 2.5 x 1012 VP
Capacity/Prep (Supernatant) One T75 flask
or one 10cm dish Four T75 flasks
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Adenoviral Expression
55
QuickTiter™ Adenoviral Titer & Quantitation Kits
QuickTiter™ Adenovirus Titer Immunoassay Kit. 293AD cells (p. 42) were infected with different dilutions of purified Ad-β-Gal for 48 hours. Immunostaining was performed according to the assay protocol. X-gal staining was performed with β-Galactosidase Stain-ing Kit (p. 106).
Accurate measurement of virus titer is critical for viral gene delivery. Traditional plaque-forming unit (PFU) assays are long and have high inter-assay variability. The QuickTiter™ Adenovirus Titer Kits provide a complete system to functionally titer virus infectivity with greater accuracy in a fraction of the time. The assays may be used with any adenovirus system that can amplify in 293 cells. Assays are available for ICC staining or 96-well ELISA. For a quick test of physical titer, our QuickTiter™ Adenovirus Quantitation Kit measures the concentra-tion of your adenovirus prep in about one hour.
Faster, More Accurate and Precise: Compared to traditional plaque-forming unit assays
User-Friendly Protocol: No agar overlay steps Versatile: Recognize all 41 adenovirus serotypes
Product Name Detection Size Catalog Number
QuickTiter™ Adenovirus Titer Immunoassay Kit ICC Staining 100 Assays VPK-109
QuickTiter™ Adenovirus Titer ELISA Kit Colorimetric 2 x 96 Assays VPK-110
QuickTiter™ Adenovirus Quantitation Kit Fluorometric 20 Assays VPK-106
Recent Product Citations 1. Smith, M. et al. (2010). PRDM1/Blimp-1 controls effector cyto-
kine production in human NK cells. J. Immunol. 185:6058-6067. (VPK-106)
2. Reiter, C.E.N. et al. (2010). Green tea polyphenol epigallocate-chin gallate reduces endothelin-1 expression and secretion in vascular endothelial cells: roles for AMP-activated protein kinase, Akt, and FOXO1. Endocrinology 151:103-114. (VPK-106)
3. Wilkins, H. et al. (2013). Mitochondrial glutathione transport is a key determinant of neuronal susceptibility to oxidative and nitrosative stress. J. Biol. Chem. 288:5091-5101. (VPK-109)
4. Scallan, C. et al. (2013). An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. 20:85-94. (VPK-109)
5. Xiong, X. et al. (2012). The autophagy-related gene 14 (Atg14) is regulated by forkhead box O transcription factors and cir-cadian rhythms and plays a critical role in hepatic autophagy and lipid metabolism. J. Biol. Chem. 287:39107-39114. (VPK-109)
6. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992. (VPK-109)
7. Hisamitsu, T. et al. (2012). Na+/H+ exchanger 1 directly binds to calcineurin A and activates downstream NFAT signaling, leading to cardiomyocyte hypertrophy. Mol. Cell Biol. 32:3265-3280. (VPK-109)
8. Lee, S. et al. (2012). Adiponectin abates diabetes-induced endothelial dysfunction by suppressing oxidative stress, adhe-sion molecules, and inflammation in type 2 diabetic mice. Am. J. Heart Circ. Physiol. 303:H106-H115. (VPK-109)
QuickTiter™ Adenovirus Titer Immunoassay Kit
QuickTiter™ Adenovirus Titer ELISA Kit
QuickTiter™ Adenovirus Quantitation Kit
Functional or Physical Titer
Functional (Infectious units)
Functional (Infectious units)
Physical (Viral particles)
Assay Time 2.5 days 2.5 days 45-60 minutes
Assay Principle Antibody-based Antibody-based Total nucleic acid content
Detection Method Immunocytochemical
staining Colorimetric (ELISA)
plate reader Fluorescence plate reader
Key Benefit Accuracy Accuracy Speed
Selection Guide for QuickTiter™ Adenoviral Quantitation Kits
www.cellbiolabs.com [email protected]
VIRAL EXPRESSION Adenoviral Expression
56
Rapid Replication Competent Adenovirus (RCA) Assay Kit
Product Name Detection Size Catalog Number
Rapid RCA Assay Kit 30 Assays VPK-111
5 x 30 Assays VPK-111-5 ICC Staining
This kit uses the assay principles of the QuickTiter™ Adenovirus Titer Immunoassay Kit (see page 47), but is designed specifically to measure the level of repli-cation-competent virus in your adenoviral prep.
Adenovirus infection of target cells is mediated largely by the coxsackievirus-adenovirus receptor (CAR). Generally adenoviral transduction of many immortalized cell lines proceeds with a high level of efficiency. However, in many primary cells this re-ceptor is either absent or present at extremely low-levels. This can reduce the efficiency of adenovirus transduction into your cell of choice. ViraDuctin™ Adenovirus Transduction Reagent is designed specifically to increase the efficiency of adenoviral transduction, without regard to the level of CAR expression on the surface of the target cells.
Higher Transduction Efficiency: Up to 12-fold increase in adenoviral uptake
User-Friendly: Short incubation step prior to host cell infection
Versatile: Ideal for target cells expressing little or no CAR, but may also improve transduction efficiency for CAR-expressing cells
Product Name Size* Catalog Number
10 Transductions AD-200
50 Transductions AD-201 ViraDuctin™ Adenovirus Transduction Reagent (CAR-Independent)
Enhanced Transduction using ViraDuctin™ Adenovirus Trans-duction Reagent. Infection of NIH3T3 cells with recombinant Ad-ß-gal (ADV-002). Top: X-gal staining under microscope. Bottom: scoring of infection with ViraDuctin™ reagent as a percentage of infection with control.
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ViraDuctin™ Adenovirus Transduction Reagent, CAR-Independent
Recent Product Citations 1. Haidar, M. et al. (2012). Integrin a2ß1 mediates tyrosine phos-
phorylation of vascular endothelial cadherin induced by inva-sive breast cancer cells. J. Biol. Chem. 287:32981-32992.
2. Ackerman, W. et al (2008). Nuclear Factor-kappa B regulates inducible prostaglandin E synthase expression in human am-nion mesenchymal cells. Biol. Reprod. 78:68-76.
3. Monick, M. et al. (2008). Constitutive ERK MAPK activity regulates macrophage ATP production and mitochondrial integrity. J. Immunol. 180:7485-7496.
*Based on using 6-well plates or 35mm culture dishes; may also be used with 96-,24- or 12-well plates or 60mm or 100mm dishes.
Faster Results: 2.5 days vs. 10 days with plaque assay
Versatile: Recognizes all 41 adenovirus serotypes
Immunostaining of Wild Type Ad5 using the Rapid RCA Assay Kit.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
57
VIRAL EXPRESSION Lentiviral Expression
Lentiviral Expression Kits & Reagents
As a sub-class of retroviruses, lentiviruses based on HIV-1 have the unique advantage of being able to infect both proliferating and non-proliferating cells, and they can be used for both transient and stable gene expression. We offer a complete workflow solution to your lentiviral expression studies:
Concentration / Purification Kits Quantitation / Titer Kits Transduction Reagents
Expression Systems & Vectors Premade Controls Viral Host Cell Line
ViraSafe™ Lentiviral Expression Systems
Lentiviruses based on HIV-1 may infect both dividing and non-dividing cells. Recently developed third-generation lentiviral expression systems have re-duced the risk of creating replication-competent virus upon recombination, but the risk is still present. Our ViraSafe™ Lentiviral Expression Systems pro-vide a safer and more flexible method to package your lentivirus, even compared to other third-generation lentivirus systems.
Safer: 80-90% less sequence homology compared to other 3rd-generation lentiviral systems; ecotropic systems provide even more safety*
High Titers: Incorporates elements that provide titers comparable to other 3rd-generation systems
Flexible: Packaging vectors provided separately for increased safety and optimization of vector ratios
Lentivirus Production using the ViraSafe™ Lentiviral Expression System.
*Lentiviruses made with a ViraSafe™ Ecotropic Expression System will only readily infect mouse and rat cells. Pan-tropic lentiviruses are VSVG-pseudotyped and may infect cells of any species.
ViraSafe™ Lentiviral Technology is available in three formats (see next two pages for ordering information): Complete Expression Systems: Include 3 packag-
ing plasmids, expression vector and control vector Packaging Systems: Include the 3 individual pack-
aging plasmids; ideal if you already have a 3rd-generation lentiviral expression construct
Expression Vectors: 11 cloning vectors to choose from; compatible with any 2nd or 3rd generation packaging system, but produce the highest titers with the ViraSafe™ packaging system
www.cellbiolabs.com [email protected]
58
VIRAL EXPRESSION Lentiviral Expression
Product Name Envelope Size Catalog Number
ViraSafe™ Universal Lentiviral Expression System (Promoterless) Ecotropic 1 Kit VPK-211-ECO
Pantropic (VSVG) 1 Kit VPK-211-PAN
ViraSafe™ Lentiviral Expression System (Puro) Ecotropic 1 Kit VPK-212-ECO
Pantropic (VSVG) 1 Kit VPK-212-PAN
ViraSafe™ Lentiviral Expression System (Neo) Ecotropic 1 Kit VPK-213-ECO
Pantropic (VSVG) 1 Kit VPK-213-PAN
ViraSafe™ Lentiviral Expression System (Hygro) Ecotropic 1 Kit VPK-214-ECO
Pantropic (VSVG) 1 Kit VPK-214-PAN
ViraSafe™ Lentiviral Bicistronic Expression System (Puro) Ecotropic 1 Kit VPK-215-ECO
Pantropic (VSVG) 1 Kit VPK-215-PAN
ViraSafe™ Lentiviral Bicistronic Expression System (Neo) Ecotropic 1 Kit VPK-216-ECO
Pantropic (VSVG) 1 Kit VPK-216-PAN
ViraSafe™ Lentiviral Bicistronic Expression System (Hygro) Ecotropic 1 Kit VPK-217-ECO
Pantropic (VSVG) 1 Kit VPK-217-PAN
ViraSafe™ Lentiviral Bicistronic Expression System (GFP) Ecotropic 1 Kit VPK-218-ECO
Pantropic (VSVG) 1 Kit VPK-218-PAN
ViraSafe™ Lentiviral Bicistronic Expression System (Blasticidin) Ecotropic 1 Kit VPK-219-ECO
Pantropic (VSVG) 1 Kit VPK-219-PAN
Ecotropic 1 Kit VPK-221-ECO
Pantropic (VSVG) 1 Kit VPK-221-PAN
ViraSafe™ shRNA Lentiviral Expression System (GFP) Ecotropic 1 Kit VPK-222-ECO
Pantropic (VSVG) 1 Kit VPK-222-PAN
ViraSafe™ shRNA Lentiviral Expression System (Puro)
ViraSafe™ Lentiviral Expression Systems, continued
Complete ViraSafe™ Expression Systems include three individual packaging plasmids, an expression vector, and a control vector. Choose an ecotropic system for infection of mouse or rat cells, or a pan-tropic system to produce VSVG-pseudotyped lenti-virus for infection of cells from any species.
ViraSafe™ Lentiviral Packaging Systems
ViraSafe™ Packaging Systems contain 3 packaging plasmids for use with any 3rd-generation lentiviral expression vector. These systems are perfect if you already have a lentiviral construct containing your gene of interest.
Product Name Envelope Size Catalog Number
ViraSafe™ Lentiviral Packaging System Ecotropic 1 Kit VPK-205
1 Kit VPK-206 Pantropic (VSVG)
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citation Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214-PAN)
Recent Product Citation Vogt, J. et al. (2014). Protein associated with SMAD1 (PAWS1/FAM83G) is a substrate for type I bone morphogenetic protein receptors and modulates bone morphogenetic protein signaling. Open Bio. 4:130210. (VPK-206)
Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalian genes induce partially reprogrammed pluripotent stem cells in non-mammalian vertebrate and
invertebrate species. eLife Sci. 2:e00036. 2. Dillahunt, S. et al. (2013). Usage of sphingosine kinase isoforms in mast cells is species and/or cell type determined. J. Immunol.
190:2058-2067.
59
VIRAL EXPRESSION Lentiviral Expression
293LTV Lentiviral Cell Line
Product Name Size Catalog Number
293LTV Cell Line 1 x 106 Cells LTV-100
ViraSafe™ Lentiviral Expression Vectors
Product Name Size Catalog Number
pSMPUW Universal Lentiviral Expression Vector (Promoterless) 10 µg VPK-211
pSMPUW-Puro Lentiviral Expression Vector 10 µg VPK-212
pSMPUW-Neo Lentiviral Expression Vector 10 µg VPK-213
pSMPUW-Hygro Lentiviral Expression Vector 10 µg VPK-214
pSMPUW-IRES-Puro Lentiviral Expression Vector 10 µg VPK-215
pSMPUW-IRES-Neo Lentiviral Expression Vector 10 µg VPK-216
pSMPUW-IRES-GFP Lentiviral Expression Vector 10 µg VPK-218
pSMPUW-IRES-Bsd Lentiviral Expression Vector 10 µg VPK-219
pSMPUW-U6-Puro Lentiviral Expression Vector 10 µg VPK-221
pSMPUW-IRES-Hygro Lentiviral Expression Vector 10 µg VPK-217
pSMPUW-U6-GFP Lentiviral Expression Vector 10 µg VPK-222
Cloning Capacity
9.4 kb
7.9 kb
7.7 kb
7.4 kb
7.8 kb
7.5 kb
7.3 kb
7.6 kb
7.9 kb
7.7 kb
7.6 kb
These lentiviral expression vectors may be used with any 2nd or 3rd generation lentiviral packaging sys-tem, but best results are achieved when used with our ViraSafe™ Lentiviral Packaging Systems.
Lentiviral Control Plasmids
Product Name Size Catalog Number
pLenti-GFP Lentiviral Control Vector 10 µg LTV-400
pSMPUW-GFP-Puro Lentiviral Control Vector 10 µg LTV-401
pSMPUW-MNDnLacZ Lentiviral Control Vector 10 µg LTV-402
pLenti-RFP-Puro Lentiviral Control Vector 100 µL LTV-403
Premade Reporter Lentivirus Controls
Product Name Concentration Size Catalog Number
GFP Lentivirus Control 1 x 106 TU/mL 200 µL LTV-300
RFP Lentivirus Control 1 x 106 TU/mL 200 µL LTV-301
www.cellbiolabs.com [email protected]
Recent Product Citations 1. Sankaran, V.G. et al. (2011). MicroRNA-15a and -16-1 act via
MYB to elevate hemoglobin expression in human trisomy 13. PNAS 108(4):1519-1524. (VPK-212)
2. Davis, M. et al. (2013). RAC1P29S is a spontaneously activating cancer-associated GTPase. PNAS 110:912-917. (VPK-214)
Our 293LTV cell line was selected from the parental 293T cell line for firmer attachment to culture plates and larger, rounder morphology for greater lentiviral production.
Assay Principle for the QuickTiter™ Lentivirus Titer Kit. Lenti-virus particles are packaged with p24 protein, but additional free p24 protein is present in viral supernatant. A traditional p24 ELISA detects both sources of p24 which overestimates viral titer. The QuickTiter™ Lentivirus Titer Kit uses technology to pull the virus out of solution prior to quantitation for a more accurate viral titer.
60
VIRAL EXPRESSION Lentiviral Expression
QuickTiter™ Lentivirus Titer / Quantitation Kits
Measuring lentiviral titer is important prior to infection of your target cells, and one of the most published methods is the p24 ELISA. Our traditional p24 ELISA kit provides a quick, convenient way to quantify the concentration of your HIV-1 based lentivirus. One disadvantage of using a traditional p24 ELISA to quantify lentivirus is the overexpression of p24 during lentiviral packaging. Free p24 protein may account for a substantial portion of total p24 in lentiviral super-natant. The traditional p24 ELISA detects both virus-associated p24 and free p24 generated by 293T cells during transient transfection. Our QuickTiter™ Lenti-virus Titer Kit minimizes the overestimation of p24 in lentivirus supernatant. Our proprietary technology separates the lentivirus-associated p24 from free p24 protein prior to performing the ELISA. If you need a very quick estimate of your lentiviral concentration, try the QuickTiter™ Lentivirus Quanti-tation Kit. This kit specifically measures the viral nu-cleic acid content of purified virus or unpurified viral supernatant. This method is ideal for a quick meas-urement of viral titer, either before or after purification of your lentivirus.
More Accurate: Exclusive technology in Quick-Titer™ Lentivirus Titer Kit minimizes overestima-tion of virus titer
User-Friendly: Read results on a standard microplate reader
Selection Guide for QuickTiter™ Lentivirus Quantitation & Titer Kits
QuickTiter™ Lentivirus Titer Kit
(Lentivirus-Associated p24 ELISA)
QuickTiter™ Lentivirus Quantitation Kits
(Traditional p24 ELISA)
QuickTiter™ Lentivirus
Quantitation Kit
Assay Principle p24 ELISA with proprietary
technology to separate free p24 from viral p24
p24 ELISA Measures
nucleic acid content
Suitable Viruses Recombinant HIV-1 Recombinant or native
HIV-1 HIV-1, FIV, SIV
Detection Method Colorimetric (ELISA)
plate reader Colorimetric (ELISA)
plate reader Fluorescence plate reader
Key Benefit Accuracy Most Published Speed (45-60 min.)
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
QuickTiter™ Lentivirus Titer / Quantitation Kits, continued
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VIRAL EXPRESSION Lentiviral Expression
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Product Name Detection Size Catalog Number
QuickTiter™ Lentivirus Titer Kit (Lentivirus-Associated HIV p24 ELISA) Colorimetric 96 Assays VPK-107
5 x 96 Assays VPK-107-5
QuickTiter™ Lentivirus Quantitation Kit (HIV-1 p24 ELISA) Colorimetric 96 Assays VPK-108-H
5 x 96 Assays VPK-108-H-5
QuickTiter™ Lentivirus Quantitation Kit Fluorometric 20 Assays VPK-112
Free p24 Does not Complex with ViraBind™ Reagents. Recom-binant p24 was diluted in culture medium and treated with Vira-Bind™ Lentivirus Reagents A and B found in the QuickTiter™ Lentivirus Titer Kit. The amount of p24 in the supernatant and the pellet was measured according to the assay protocol.
Recent Product Citations 1. Belaner, K. et al. (2013). Binding of RNA by APOBEC3G con-
trols deamination-independent restriction of retroviruses. J. Exp. Biol. 216:2213-2220. (VPK-107)
2. Yu, X. et al. (2012). Identification of Hepatitis B virus inhibitors targeting different aspects of infection using a cell-based assay. Antimicrob. Agents Chemother. 56:6109-6120. (VPK-107)
3. Walker, K. et al. (2012). Depletion of GGA1 and GGA3 mediates postinjury elevation of BACE1. J. Neurosci. 32:10423-10437. (VPK-107)
4. Zhou, B. et al. (2012). Interactions between ß-catenin and Transforming growth factor-ß signaling pathways mediate epithelial-mesenchymal transition and are dependent on the transcriptional co-activator cAMP-response element-binding protein (CREB)-binding. J. Biol. Chem. 287:7026-7038. (VPK-107)
5. Nedelec, A.D. et al. (2012). Noonan Syndrome-causing SHP2 mutants inhibit insulin-like growth factor 1 release via growth hormone-induced ERK hyperactivation, which contributes to short stature. PNAS 109:4257-4262. (VPK-107)
6. Lavender, H. et al. (2012). In vitro characterization of the activity of PF-05095808, a novel biological agent for Hepatitis C virus therapy. Antimicrob. Agents Chemother. 56:1364-1375. (VPK-107)
7. Keck, Z.Y. et al. (2011). Mapping a region of Hepatitis C virus E2 that is responsible for escape from neutralizing antibodies and a core CD81-binding region that does not tolerate neutrali-zation escape mutations. J. Virol. 85:10451-10463. (VPK-107)
8. Sanchez-Antequera, Y. et al. (2011). Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells. Blood 117:e171-e181. (VPK-107)
9. Yi, S.H. et al. (2014). Foxa2 acts as a co-activator potentiating expression of the Nurr1-induced DA phenotype via epigenetic regulation. Development 141:761-772. (VPK-108-H)
10. Smith, B. et al. (2013). Targeting the PyMT oncogene to diverse mammary cell populations enhances tumor heterogeneity and generates rare breast cancer subtypes. Genes & Cancer 10.1177/1947601913475359. (VPK-108-H)
11. Iftikhar, M. et al. (2011). Lysyl oxidase-like-2 (LOXL2) is a major isoform in chondrocytes and is critically required for differentia-tion. J. Biol. Chem. 286:909-918. (VPK-108-H)
12. Agrawal-Gamse, C. et al. (2010). Yeast-elicited cross-reactive to HIV Env glycans efficiently neutralize virions expressing exclu-sively high mannose N-linked glycans. J. Virol. 85(1):470-480. (VPK-108-H)
13. Rossello, R.A. et al. (2013). Mammalina genes induce partially reprogrammed pluripotent stem cells in non-mammalian verte-brate and invertebrate species. eLife Sci. 2:e00036. (VPK-112)
14. Fan, X. et al. (2012). Transient, inducible, placenta-specific gene expression in mice. Endocrinology 153:5637-5644. (VPK-112)
15. Veeraraghavalu, K. et al. (2010). Presinilin 1 mutants impair the self-renewal and differentiation of adult murine subventricular zone-neuronal progenitors via cell-autonomous mechanisms involving notch signaling. J. Neurosci. 30:6903-6915. (VPK-112)
www.cellbiolabs.com [email protected]
VIRAL EXPRESSION Lentiviral Expression
62
ViraBind™ Lentivirus Concentration & Purification Kits
Lentivirus Concentration and Purification Procedure.
Ultracentrifugation methods used for lentiviral super-natants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filter-based methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Lentivirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isola-tion technology, then resuspended in a smaller vol-ume. The resuspended lentivirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be fur-ther concentrated using a centrifugal concentrator.
Fast: Obtain purified virus in about 4-6 hours with column-based kits and 10-24 hours with dialysis-based kits
High Titer: Concentrate up to 500-fold to as high as 108-1010 TU/ml, sufficient for in vivo studies
High Yield: Recover >60%
Product Name Size Catalog Number
ViraBind™ Lentivirus Concentration and Purification Kit (100 ml/prep)
2 Preps VPK-090
5 Preps VPK-091
25 Preps VPK-091-5
ViraBind™ PLUS Lentivirus Concentration and Purification Kit (50 ml/prep) 2 Preps VPK-095
2 Preps VPK-096
10 Preps VPK-096-5 ViraBind™ PLUS Lentivirus Concentration and Purification Mega Kit (500 ml/prep)
Selection Guide for Lentivirus Concentration & Purification Kits
ViraBind™ Lentivirus
Concentration and Purification Kit
ViraBind™ PLUS Lentivirus
Concentration and Purification Kit
ViraBind™ PLUS Lentivirus
Concentration and Purification Mega Kit
Purification Method
Proprietary Reagent Cocktail + Purification Column
Proprietary Reagent Cocktail + Dialysis
Proprietary Reagent Cocktail + Dialysis
Total Time 6-8 hours 10-24 hours 10-24 hours
Capacity per Prep(Supernatant)
100 mL 50 mL 500 mL
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Lentiviral Expression
63
ViraDuctin™ Lentivirus Transduction Kit
Product Name Size* Catalog Number
40 Transductions LTV-200
200 Transductions LTV-201 ViraDuctin™ Lentivirus Transduction Kit
Transduction of 293AD and HT-1080 Cells. 293AD cells (p. 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24-well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of no additive (left), Polybrene® (middle) or the ViraDuctin™ reagent cocktail (right).
Transduction Efficiencies in Various Cell Lines. NIH3T3 cells, HeLa cells, our own 293AD cells (page 36) and HT-1080 cells were each seeded at 50,000 cells/well in a 24-well plate overnight. Cells were infected with GFP lentivirus for 48 hours in the presence of Polybrene® or ViraDuctin™ Lentivirus Transduc-tion Kit. For each cell line, fluorescence levels using the Vira-Ductin™ system are depicted relative to a normalized fluores-cence of 100 for Polybrene®.
Lentivirus transduction efficiency is typically low. Ad-ditives such as Polybrene® can boost transduction efficiencies, but even then only a small fraction of len-tiviral vectors can transduce many target cell lines. Our ViraDuctin™ Lentivirus Transduction Kit provides superior transduction efficiencies in a variety of cell lines, even when compared to transductions in the presence of Polybrene®.
Higher Transduction Efficiency: 2-6x higher in many cell lines compared to Polybrene
More Robust: Useful for transduction of nonper-missive cells, including primary cells and stem cells
Polybrene is a registered trademark of Abbott Laboratories.
*Based on a 24-well plate. Can also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert.
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www.cellbiolabs.com [email protected]
Recent Product Citations 1. Rossello, R.A. et al. (2013). Mammalina genes induce partially
reprogrammed pluripotent stem cells in non-mammalian verte-brate and invertebrate species. eLife Sci. 2:e00036.
2. McEachron, T.A. et al. (2010). Protease-activated receptors mediate crosstalk between coagulation and fibrinolysis. Blood 116:5037-5044.
3. Zemskova, M. et al. (2010). p53-dependent induction of prostate cancer cell senescence by the PIM1 protein kinase. Mol. Cancer Res. 8:1126-1141.
VIRAL EXPRESSION Retroviral Expression
64
Retroviral Expression Kits & Reagents
Traditional retroviral vectors based on MMLV are useful for integrating genetic material into the host cell genome. However, retrovirus titer tends to be significantly lower than that of adenovirus, which can lead to a lower infection efficiency. Our retroviral reagents and kits incorporate technologies that increase your chances of successful retroviral expression. We offer a comprehensive solution from start to finish:
Gene-Specific Retroviral Vectors Concentration / Purification Kits Quantitation / Titer Kits Transduction Reagents
Retroviral Expression Systems Retroviral Packaging Cell Lines Retroviral Cloning & Expression
Vectors
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Product Name Size Catalog Number
Platinum-E Retroviral Packaging Cell Line, Ecotropic >3 x 106 cells RV-101
Platinum-A Retroviral Packaging Cell Line, Amphotropic >3 x 106 cells RV-102
Platinum-GP Retroviral Packaging Cell Line, Pantropic >3 x 106 cells RV-103
pVSV-G Packaging Vector 10 µg RV-110
Platinum Retroviral Packaging Cell Lines
Recent Product Citations 1. Schmidt, T. et al. (2013). CXCR4 promotes B cell egress from
Peyer’s patches. J. Exp. Med. 10.1084/jem.20122574. (RV-101) 2. Wahlestedt, M. et al. (2013). An epigenetic component of hemato-
poietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RV-101)
3. Zhong, S. et al. (2013). T-cell receptor affinity and avidity defines antitumor response and autoimmunity in T-cell immunotherapy. PNAS 110:6973-6978. (RV-101)
4. Nam, Y.J. et al. (2013). Reprogramming of human fibroblasts to-ward a cardiac fate. PNAS 110:5588-5593. (RV-102)
5. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase re-verse transcriptase variants lacking telomerase activity stimulate cell proliferation. Mol. Cell Biol. 32:4283-4296. (RV-102)
6. Nowakowski, T. et al. (2013). MicroRNA-92b regulates the devel-opment of intermediate cortical progenitors in embryonic mouse brain. PNAS 110:7056-7061. (RV-103)
7. Cavnar, P.J. et al. (2012). The actin regulatory protein HS1 inter-acts with Arp2/3 and mediates efficient neutrophil chemotaxis. J. Biol. Chem. 287:25466-25477. (RV-103)
Generate high titers of recombinant retrovirus with a single plasmid transfection* using these extremely powerful, stable cell lines. Platinum Retroviral Packag-ing Cells are based on the 293T cell line and exhibit greater stability and produce higher yields of retroviral structure proteins, resulting in higher retroviral titers. The Platinum cell lines were invented in the laboratory of Dr. Toshio Kitamura at the University of Tokyo and are available exclusively from Cell Biolabs. They were first described in the following paper: Morita, S. et al. (2000). Gene Therapy 7:1063-1066.
Plat-A Cells(Amphotropic)
Plat-E Cells(Ecotropic)
Plat-GP Cells(Pantropic*)
Human +++ N.S. +++
Mouse +++ +++ +++
Rat +++ +++ +++
Monkey +++ N.S. +++
Cat +++ N.S. +++
Dog +++ N.S. +++
Hamster + N.S. +++
Bird N.S. N.S. +++
Fish N.S. N.S. +++
Frog N.S. N.S. +++
Insect N.S. N.S. +++
Mollusk N.S. N.S. +++
*Plat-GP cells must be co-transfected with a pantropic envelope protein such as VSV-G. N.S. = Not Suitable
Suitability of Platinum Retroviral Packaging Cell Lines by Host Species.
Not sure which Platinum Expression System is right for you? See the table below for a selection guide based on the host species of your target cell.
VIRAL EXPRESSION Retroviral Expression
65 www.cellbiolabs.com [email protected]
Platinum Retroviral Packaging Cells and Expression Systems
Our Platinum Retroviral Expression Systems incorpo-rate superior packaging cell lines and vector technolo-gies to produce high-titer virus with a single plasmid transfection. Each Platinum Expression System in-cludes one of our exclusive Platinum Packaging Cell Lines which stably express the gag and pol genes. In the Ecotropic and Amphotropic systems, the packag-ing cells also express the envelope protein.* Simply clone your gene of interest into the vector provided and transfect into the Platinum cells. Platinum Retroviral Expression Systems contain eve-rything you need to generate your recombinant retro-virus: packaging cell line, expression vector, and GFP control vector. Our pantropic systems also contain a VSVG envelope vector.
Retrovirus Production Using the Platinum Expression Systems (Ecotropic and Amphotropic).
Higher Viral Yields: Average titer 107 infectious units/mL with transient transfection
Longer Stability: Expression up to 4 months in the presence of drug selection
Optimized Systems: 3 packaging cell lines for in-fection of various species; 3 vector backbones (two specifically for infection of stem cells)
Flexible: Order complete systems or cells and vec-tors separately
Product Name Expression Vector Packaging Cell Catalog Number
Platinum Retroviral Expression System, Ecotropic pMXs-Puro Plat-E VPK-300
Platinum Retroviral Expression System, Amphotropic pMXs-Puro Plat-A VPK-301
Platinum Retroviral Expression System, Pantropic pMXs-Puro Plat-GP VPK-302
Platinum ES/EC Retroviral Expression System, Ecotropic pMCs-Puro Plat-E VPK-303
Platinum ES/EC Retroviral Expression System, Amphotropic pMCs-Puro Plat-A VPK-304
Platinum ES/EC Retroviral Expression System, Pantropic pMCs-Puro Plat-GP VPK-305
Platinum HSC Retroviral Expression System, Ecotropic pMYs-Puro Plat-E VPK-306
Platinum HSC Retroviral Expression System, Amphotropic pMYs-Puro Plat-A VPK-307
Platinum HSC Retroviral Expression System, Pantropic pMYs-Puro Plat-GP VPK-308
Recent Product Citations 1. Aoi, N. et al. (2012). 1a,25-dihydroxyvitamin D3 modulates the hair
-inductive capacity of dermal papilla cells: therapeutic potential for hair regeneration. Stem Cells Trans Med. 1:615-626. (VPK-301)
2. Wang, N. et al. (2013). Lacritin rescues stressed epithelia via rapid forkhead box O3 (FOXO3)-associated autophagy that restores metabolism. J. Biol. Chem. 288:18146-18161. (VPK-302)
3. Tanaka, T. et al. (2012). Anthracycline inhibits recruitment of hy-poxia-inducible transcription factors and suppresses tumor cell migration and cardiac angiogenic response in the host. J. Biol. Chem. 287:34866-34882. (VPK-302)
*Pantropic systems require co-transfection with the provided VSVG envelope vector.
66
VIRAL EXPRESSION Retroviral Expression
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Retroviral Cloning & Expression Vectors
Our Retroviral Expression Vectors are based on backbones derived from Moloney murine leukemia virus (MMLV). We offer the traditional pBABE system and the novel pMXs system, which has been shown to be useful in induced pluripotent stem cell (iPS) studies. pMYs vectors are optimal for use with hema-topoietic stem cells, and pMCs vectors are optimal for ES and EC cells. All cloning vectors are supplied as 10 µg in TE buffer.
Vector Name Cloning Capacity Catalog Number
pMXs-U6-GFP 5 kb RTV-071
pMXs-U6-Puro 5.1 kb RTV-070
pMXs-U6-Puro-shGFP RTV-055
pMXs-U6-Puro-shLuc RTV-056
Recent Product Citations 1. Duran, P.P. et al. (2012). UNG shapes the specificity of AID-
induced somatic hypermutation. J. Exp. Med. 209:1379-1389. (RTV-001-HYGRO)
2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45. (RTV-010)
3. Parikh, C. et al. (2012). Disruption of PH-kinase domain interac-tions leads to oncogenic activation of AKT in human cancers. PNAS 109:19368-19373. (RTV-012)
4. Zhang, Q. et al. (2013). TNF-a impairs differentiation and func-tion of TGF-ß-induced Treg cells in autoimmune diseases through Akr and Smad3 signaling pathway. J. Mol. Cell Biol. 10.1093/jmcb/jms063. (RTV-013)
5. Sugatani, T. et al. (2011). A microRNA expression signature of osteoclastogenesis. Blood 117:3648-3657. (RTV-014, RTV-016)
Retroviral Cloning Vectors for General Gene Expression (driven by 5’ LTR)
Vector Name Cloning Capacity Catalog Number
pBABEhygro 5.6 kb RTV-001-HYGRO
pBABEneo 5.9 kb RTV-003
pBABEpuro 6 kb RTV-001-PURO
pBABEzeo 6.3 kb RTV-004
pMXs 5.4 kb RTV-010
pMXs-IRES-Bsd 5.6 kb RTV-016
pMXs-IRES-GFP 5.3 kb RTV-013
pMXs-IRES-Neo 5.2 kb RTV-015
pMXs-IRES-Puro 5.4 kb RTV-014
pMXs-Neo 3.8 kb RTV-011
pMXs-Puro 4.4 kb RTV-012
pMZs 5.3 kb RTV-030
Retroviral Cloning Vectors with Strong Promoters for Overexpression
Vector Name Cloning Capacity Catalog Number
pMXs-CAG 5.2 kb RTV-064
pMXs-CMV 5.5 kb RTV-065
pMXs-EF1 5.5 kb RTV-063
pMXs-EF1-Bsd 4.2 kb RTV-062
pMXs-EF1-GFP 3.9 kb RTV-061
pMXs-EF1-Puro 4 kb RTV-060
pMXs-SR 5.4 kb RTV-066
Retroviral Cloning Vectors for use with ES/EC Cells
Vector Name Cloning Capacity Catalog Number
pMCs-IRES-GFP 5.2 kb RTV-040
pMCs-Puro 4.3 kb RTV-041
Retroviral Cloning Vectors for use with Hematopoietic Cells
Vector Name Cloning Capacity Catalog Number
pMYs 5.2 kb RTV-020
pMYs-IRES-GFP 5.2 kb RTV-021
pMYs-IRES-Neo 5.2 kb RTV-023
pMYs-IRES-Puro 5.4 kb RTV-022
pMYs-Puro 4.3 kb RTV-024
Retroviral Cloning Vector for miRNA
Vector Name Cloning Capacity Catalog Number
pMXs-miR-GFP/Puro 4.2 kb RTV-017
Retroviral Cloning Vectors for shRNA
Recent Product Citations 1. Malicet, C. et al. (2011). Distinct properties of human HMGN5
reveal a rapidly evolving but functionally conserved nucleosome binding protein. Mol. Cell Biol. 31:2742-2755. (RTV-040)
2. Mochizunki, Y. et al. (2013). Phosphatidylinositol 3-phosphate myotubularin-related protein 6 (MTMR6) is regulated by small GTPase Rab1b in the early secretory and autophagic pathways. J. Biol. Chem. 288:1009-1021. (RTV-041)
Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017)
VIRAL EXPRESSION Retroviral Expression
67 www.cellbiolabs.com
Retroviral Packaging Vectors and Cells
These constructs are ideal for researchers who prefer a traditional multi-plasmid transfection of 293 cells for packaging recombinant retrovirus.
Product Name Size Catalog Number
pCMV-10A1 Envelope Vector 100 µL RV-114
pCMV-Ampho Envelope Vector 100 µL RV-113
pCMV-Eco Envelope Vector 100 µL RV-112
pCMV-Gag-Pol Retroviral Vector 10 µg RV-111
pCMV-VSV-G Envelope Vector 10 µg RV-110
Recent Product Citations 1. Amagai, Y. et al. (2013). Stem cell factor contributes to tumorigenesis of mast cells via an autocrine/paracrine mechanism. J. Leukoc.
Biol. 93:245-250. (RV-110) 2. Okamoto, K. et al. (2012). Dengue virus strain DEN2 16681 utilizes a specific glycochain of syndecan-2 proteoglycan as a receptor. J.
Gen. Virol. 93:761-770. (RV-110, RV-111)
Target Name Vector Backbone Catalog Number
c-Abl pBABEpuro RTV-402
c-Abl-TM pBABEpuro RTV-403
c-Abl (1-565) pBABEpuro RTV-404
c-Abl (1-958) pBABEpuro RTV-405
pBABEhygro RTV-007
pBABEneo RTV-005
pBABEpuro RTV-006
p53 pBABEpuro RTV-401
hTERT
Cell Cycle
Recent Product Citation Huang, J. et al. (2009). Regulation of the leucocyte chemoattrac-tant receptor FPR in glioblastoma cells by cell differentiation. Carcinogenesis 30(2):348-355. (RTV-401)
Reporter Genes
Target Name Catalog Number
GFP RTV-002
GFP RTV-051
Vector Backbone
pBABE
pMCs
GFP pMX RTV-050
GFP pMYs RTV-052
GFP-Puro pMX RTV-053
Recent Product Citations 1. Hrdlickova, R. et al. (2012). Alternatively spliced telomerase
reverse transcriptase variants lacking telomerase activity stimu-late cell proliferation. Mol. Cell Biol. 32:4283-4296. (RTV-002)
2. Wahlestedt, M. et al. (2013). An epigenetic component of hema-topoietic stem cell aging amenable to reprogramming into a young state. Blood 121:4257-4264. (RTV-050)
Autophagy
Target Name Vector Backbone Catalog Number
GFP-LC3 pMXs RTV-801
This vector is supplied with a separate pMXs-GFP control vector at no additional cost.
These constructs are based on backbones derived from MMLV, Vectors with GFP or stem cell factors are supplied as 10 µg of plasmid in TE buffer. All other vec-tors are supplied as 100 µL of bacterial glycerol stock. Product listing continues on the following pages.
Gene-Specific Recombinant Retroviral Vectors
293RTV Cell Line
Product Name Size Catalog Number
293RTV Cell Line >1 x 106 cells RV-100
Our 293RTV cells are derived from the 293 parental cell line, but are selected for firmer attachment to culture plates, faster growth and higher yields of retrovirus produced.
Vector Name Vector Backbone Mutation State Catalog Number
ERK2 pBABEhygro Dominant Negative RTV-109
JNK1 pBABEpuro Dominant Negative RTV-110
MAPKAPK2 pBABEpuro Constitutively Active RTV-118
pBABEpuro Dominant Negative RTV-119
MAPKAPK3 pBABEpuro Constitutively Active RTV-120
pBABEpuro Dominant Negative RTV-121
MEK1 pBABEhygro Constitutively Active RTV-112
pBABEhygro Dominant Negative RTV-111
MKK3 pBABEpuro Constitutively Active RTV-114
pBABEhygro Dominant Negative RTV-115
MKK6 pBABEpuro Constitutively Active RTV-116
pBABEhygro Dominant Negative RTV-117
myr-Akt1 pWZLneo Constitutively Active RTV-125
p38 pBABEhygro Dominant Negative RTV-105
p38 pBABEhygro Dominant Negative RTV-106
p38 pBABEhygro Dominant Negative RTV-107
p38 pBABEhygro Dominant Negative RTV-108
PI3K p110-CAAX pWZLneo Constitutively Active RTV-124
pBABEpuro Constitutively Active RTV-122
pBABEpuro Dominant Negative RTV-123
Raf1-CAAX pWZLneo Constitutively Active RTV-113
PRAK
MAP Kinase Signaling
68
VIRAL EXPRESSION Retroviral Expression
Target Name Vector Backbone Catalog Number
AUF1 pBABEpuro RTV-305
hnRNPA0 pBABEpuro RTV-310
hnRNP-A2 pBABEpuro RTV-340
HuB pBABEpuro RTV-302
HuC pBABEpuro RTV-303
HuD pBABEpuro RTV-301
HuR pBABEpuro RTV-304
PABP pBABEpuro RTV-307
Stat5A pMXs RTV-330
Stat5A(1*6) pMXs RTV-331
Transcription Regulation
Gene-Specific Recombinant Retroviral Vectors, continued
Target Name Vector Backbone Catalog Number
Stat5A-IRES-GFP pMXs RTV-332
Stat5A(1*6)-IRES-GFP
pMXs RTV-333
Stat5B pMXs RTV-334
Stat5B(1*6) pMXs RTV-335
TIA-1 pBABEpuro RTV-309
TIAR pBABEpuro RTV-308
TTP pBABEpuro RTV-306
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citation Yu, Y. et al. (2012). Bcl11a is essential for lymphoid development and negatively regulates p53. J. Exp. Med. 209:2467-2483. (RTV-331)
VIRAL EXPRESSION Retroviral Expression
69 www.cellbiolabs.com [email protected]
Gene-Specific Recombinant Retroviral Vectors, continued
Cytoskeleton Regulation
iPS / Stem Cell Factors
Target Name Vector Backbone Catalog Number
4-Vector Set* pMXs RTV-701-C
6-Vector Set** pMXs RTV-709-C
c-Myc pMXs RTV-703
Klf4 pMXs RTV-704
Lin-28 pMXs RTV-710
NANOG pMXs RTV-709
Oct-3/4 pMXs RTV-701
Sox2 pMXs RTV-702
p53 shRNA pRetro RTV-410
*4-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4 and Sox2. **6-Vector sets contain individual constructs with the following genes: c-Myc, Klf4, Oct-3/4, Sox2, Lin-28 and NANOG.
Target Name Vector Backbone Catalog Number
4-Vector Set* pMXs RTV-705-C
6-Vector Set** pMXs RTV-711-C
c-Myc pMXs RTV-707
Klf4 pMXs RTV-708
Lin-28 pMXs RTV-712
NANOG pMXs RTV-711
Oct-3/4 pMXs RTV-705
Sox2 pMXs RTV-706
p53 shRNA pRetro RTV-400
Human iPS Genes Mouse iPS Genes
Proteases and Related Molecules
Recent Product Citation Gutova, M. et al (2008). Urokinase plasminogen activator and urokinase plasminogen activator receptor mediate human stem cell tropism to malignant solid tumors. Stem Cells 26:1406-1413. (RTV-501, RTV-502)
Target Name Catalog Number
uPA RTV-501
uPAR RTV-502
Vector Backbone
pBABEpuro
pBABEhygro
Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:2467-2483. (RTV-101)
Target Name Vector Backbone Mutation State Catalog Number
Cdc42 pBABEhygro L61 RTV-203
K-Ras pBABEpuro N/A RTV-220
pWZLhygro Q61 RTV-221
myr-Rac1 N/A RTV-201
V12 RTV-206
Rac1 pBABEhygro V12 RTV-202
N-Ras pBABEpuro K61 RTV-222
Rac3 pBABEhygro V12 RTV-205
Ras
pBABEpuro V12 RTV-101
pBABEpuro V12C40 RTV-104
pBABEpuro V12G37 RTV-103
pBABEpuro V12S35 RTV-102
RhoA pBABEhygro L63 RTV-204
pBABEpuro
VIRAL EXPRESSION Retroviral Expression
70
ViraBind™ Retrovirus Concentration & Purification Kits
Fast: Obtain purified virus in about 4-6 hours High Titer: Concentrate 500-fold to 109-1010
TU/ml, sufficient for in vivo studies High Yield: Recover >60% High Throughput: Process greater volumes
per prep than filter-based purification methods
Retrovirus Concentration and Purification Procedure.
Product Name Size Catalog Number
ViraBind™ Retrovirus Concentration and Purification Kit (100 ml/prep)
2 Preps VPK-130
5 Preps VPK-131
25 Preps VPK-131-5
ViraBind™ PLUS Retrovirus Concentration and Purification Kit (50 ml/prep) 2 Preps VPK-135
2 Preps VPK-136
10 Preps VPK-136-5 ViraBind™ PLUS Retrovirus Concentration and Purification Mega Kit (500 ml/prep)
Selection Guide for Retrovirus Concentration & Purification Kits
ViraBind™ Retrovirus
Concentration and Purification Kit
ViraBind™ PLUS Retrovirus
Concentration and Purification Kit
ViraBind™ PLUS Retrovirus
Concentration and Purification Mega Kit
Purification Method
Proprietary Reagent Cocktail + Purification Column
Proprietary Reagent Cocktail + Dialysis
Proprietary Reagent Cocktail + Dialysis
Total Time 6-8 hours 10-24 hours 10-24 hours
Capacity per Prep(Supernatant)
100 mL 50 mL 500 mL
Ultracentrifugation methods used for lentiviral super-natants are tedious and time-consuming and usually only partially purify your virus. Alternatively, filter-based methods have low sample capacity and do not substantially concentrate the virus. ViraBind™ Retrovirus Concentration and Purification Kits produce purified lentivirus with extremely high titer without the need for ultracentrifugation. The virus is pelleted from solution using our proprietary isola-tion technology, then resuspended in a smaller vol-ume. The resuspended retrovirus is then applied to either a purification column or a dialysis device for purification. If desired, the purified virus may be fur-ther concentrated using a centrifugal concentrator.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
VIRAL EXPRESSION Retroviral Expression
71
QuickTiter™ Retrovirus Rapid Quantitation Kit
Product Name Detection Size Catalog Number
QuickTiter™ Retrovirus Quantitation Kit Fluorometric 20 Assays VPK-120
Ultra-fast Results: 45-60 minute procedure Convenient: Titer may be measured before purifi-
cation step Sensitive: Limit of detection = 1.5 x 109 VP/mL
from 2 mL of retroviral supernatant
This kit specifically measures the viral nucleic acid con-tent of purified virus or unpurified viral supernatant. This method is ideal for a quick measurement of viral titer, either before or after purification of your retrovirus.
0
100
200
300
400
500
0 400 800 1200
Retroviral RNA (ng)
RF
U (
52
0 n
m)
0
10
20
30
40
50
60
70
80
0 50 100 150
Retroviral RNA (ng)
RF
U (
52
0 n
m)
Retrovirus RNA Standard Curve. The QuickTiter™ Retrovirus RNA Standard was diluted according to the assay protocol. Fluo-rescence was measured on a SpectraMax Gemini XS Fluorometer (Molecular Devices) with a 485 / 538 nm filter set and a 530 nm cutoff.
Assay Procedure for the QuickTiter™ Retrovirus Quantitation Kit.
www.cellbiolabs.com [email protected]
Recent Product Citation Ito, T. et al. (2012). Stem cell factor programs the mast cell activa-tion phenotype. J. Immunol. 188:5428-5437.
VIRAL EXPRESSION Retroviral Expression
72
ViraDuctin™ Retrovirus Transduction Kit
Product Name Size* Catalog Number
40 Transductions RV-200
200 Transductions RV-201 ViraDuctin™ Retrovirus Transduction Kit
The efficiency of retrovirus transduction can be low compared to other viruses. The rate at which retrovi-ral vectors bind to cells is controlled mostly by diffu-sion. Additionally, the presence of transduction inhibi-tors such as proteoglycans and glycosaminoglycans in retroviral supernatants can lead to poor gene trans-fer. Additives such as Polybrene® can boost transduc-tion efficiencies, but they do not eliminate these trans-duction inhibitors. Our ViraDuctin™ Retrovirus Transduction Kit pro-vides superior transduction efficiencies even when compared to transductions in the presence of Poly-brene®. A proprietary reagent cocktail forms a super-complex with the retrovirus which is pelleted away from the supernatant, removing detrimental transduc-tion inhibitors that decrease infection efficiency.
More Robust: Removes harmful transduction inhibitors from retroviral supernatant
Higher Transduction Efficiencies: Compared to infections in the presence of Polybrene or no additives
Versatile: Particularly useful for nonpermissive cells including primary cells and stem cells, but may boost transduction rates in a wide variety of cells
Polybrene is a registered trademark of Abbott Laboratories.
*Number of transductions shown is based on use in a 24-well plate. This product may also be used with 96-well, 12-well or 6-well plates, as well as 60mm or 100mm dishes. See product insert for specific details.
Recent Product Citations 1. Gandhi, M. et al. (2012). Homologous chromosomes make con-
tact at the sites of double-strand breaks in genes in somatic G0/G1-phase human cells. PNAS 109:9454-9459.
2. Miyoshi, N. et al. (2010). Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS 107:40-45.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
74
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors)
Precursor Name Catalog Number
mmu-let-7a-1 MMU-LET7A-1
hsa-let-7a-2 MIR-LET7A-2
hsa-let-7a-3 MIR-LET7A-3
mmu-let-7b MMU-LET7B
hsa-let7c MIR-LET7C
mmu-let-7c-1 MMU-LET7C-1
mmu-let-7c-2 MMU-LET7C-2
hsa-let-7d MIR-LET7D
mmu-let-7d MMU-LET7D
hsa-let-7e MIR-LET7E
hsa-let-7f-1 MIR-LET7F-1
mmu-let-7f-1 MMU-LET7F-1
mmu-let-7f-2 MMU-LET7F-2
hsa-let-7g MIR-LET7G
mmu-let-7g MMU-LET7G
hsa-let-7i MIR-LET7I
hsa-mir-1-1 MIR-1-1
mmu-mir-1-1 MMU-MIR-1-1
hsa-mir-1-2 MIR-1-2
mmu-mir-1-2 MMU-MIR-1-2
hsa-mir-7-1 MIR-7-1
hsa-mir-7-2 MIR-7-2
hsa-mir-7-3 MIR-7-3
mmu-mir-7a-1 MMU-MIR-7A-1
mmu-mir-7a-2 MMU-MIR-7A-2
mmu-mir-7b MMU-MIR-7B
hsa-mir-9-1 MIR-9-1
mmu-mir-9-1 MMU-MIR-9-1
hsa-mir-9-2 MIR-9-2
hsa-mir-10a MIR-10A
hsa-mir-10b MIR-10B
mmu-mir-10b MMU-MIR-10B
hsa-mir-15a MIR-15A
mmu-mir-15a MMU-MIR-15A
mmu-mir-15b MMU-MIR-15B
miRNASelect™ Expression Vectors contain full miRNA precursor se-quences with constitutive promoter. Human (hsa) vectors contain a
puromycin selection marker Mouse (mmu) vectors contain a
GFP-puromycin fusion
Precursor Name Catalog Number
hsa-mir-17 MIR-17
hsa-mir-18a MIR-18A
mmu-mir-18a MMU-MIR-18A
hsa-mir-18b MIR-18B
mmu-mir-18b MMU-MIR-18B
hsa-mir-19a MIR-19A
mmu-mir-19a MMU-MIR-19A
hsa-mir-19b-1 MIR-19B-1
mmu-mir-19b-1 MMU-MIR-19B-1
hsa-mir-19b-2 MIR-19B-2
mmu-mir-19b-2 MMU-MIR-19B-2
hsa-mir-20a MIR-20A
mmu-mir-20a MMU-MIR-20A
hsa-mir-20b MIR-20B
mmu-mir-20b MMU-MIR-20B
hsa-mir-21 MIR-21
mmu-mir-21 MMU-MIR-21
hsa-mir-22 MIR-22
hsa-mir-23b MIR-23B
mmu-mir-23b MMU-MIR-23B
mmu-mir-24-1 MMU-MIR-24-1
hsa-mir-24-2 MIR-24-2
mmu-mir-24-2 MMU-MIR-24-2
mmu-mir-25 MMU-MIR-25
hsa-mir-26a-1 MIR-26A-1
mmu-mir-26a-1 MMU-MIR-26A-1
hsa-mir-26a-2 MIR-26A-2
mmu-mir-26a-2 MMU-MIR-26A-2
hsa-mir-26b MIR-26B
hsa-mir-27a MIR-27A
hsa-mir-27b MIR-27B
mmu-mir-27b MMU-MIR-27B
hsa-mir-16-1 MIR-16-1
mmu-mir-16-1 MMU-MIR-16-1
hsa-mir-16-2 MIR-16-2
Don’t see your microRNA of interest? Clone your own sequence into one of our
Expression Vectors; see page 80.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b
in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442. (MIR-18B)
2. Mallory, M.J. et al. (2011). Signal– and developmental-dependent alternative splicing of LEF1 in T cells is controlled by CELF2. Mol. Cell. Biol. 31:2184-2195. (MIR-23B)
3. Woo, H. et al. (2013). Nucleolin mediates microRNA-directed CSF-1 mRNA deade-nylation but increases translation of CSF-1 mRNA. Mol. Cell Proteomics 12:1661-1667. (MIR-130A, MIR-301A)
4. Starega-Roslan, J. et al. (2010). Stuctural basis of microRNA length variety. Nuc. Acids Res. 10.1093/nar/gkq727. (MIR-148A)
5. Saus, E. et al. (2010). Genetic variants and abnormal processing of pre-miR-182, a circadian clock modulator, in major de-pression patients with late insomnia. Hum. Mol. Genet. 19:4017. (MIR-182)
6. Beezhold, K. et al. (2011). miR-190-mediated downregulation of PHLPP con-tributes to arsenic-induced Akt activation and carcinogenesis. Toxicol. Sci. 123:411-420. (MIR-190)
7. Wang, Y.S. et al. (2012). MicroRNA-195 regulates vascular smooth muscle cell phenotype and prevents neointimal forma-tion. Cardiovasc. Res. 95:5174-526. (MIR-195)
8. Majid, S. et al. (2011). MicroRNA-205 inhibits src-mediated oncogenic pathways in renal cancer. Cancer Res. 71:2611-2621. (MIR-205)
9. Halappanaver, S. et al. (2013). IL-1 recep-tor regulates microRNA-135b expression in a negative feedback mechanism. J. Immunol. 190:3679-3686. (MMU-MIR-135B)
10.Yamamoto, H. et al. (2012). MicroRNA-494 regulates mitochondrial biogenesis in skeletal muscle through mitochondrial transcription factor A and forkhead box j3. Am J. Physiol. Endocrinol. Metab. 303:E1419-1427. (MMU-MIR-494)
75
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
Precursor Name Catalog Number
mmu-mir-29a MMU-MIR-29A
hsa-mir-29b-1 MIR-29B-1
mmu-mir-29b-1 MMU-MIR-29B-1
mmu-mir-29b-2 MMU-MIR-29B-2
hsa-mir-29c MIR-29C
mmu-mir-29c MMU-MIR-29C
hsa-mir-30a MIR-30A
mmu-mir-30a MMU-MIR-30A
hsa-mir-30b MIR-30B
hsa-mir-30c-1 MIR-30C-1
mmu-mir-30c-1 MMU-MIR-30C-1
hsa-mir-30c-2 MIR-30C-2
mmu-mir-30c-2 MMU-MIR-30C-2
hsa-mir-30d MIR-30D
mmu-mir-30d MMU-MIR-30D
hsa-mir-30e MIR-30E
mmu-mir-30e MMU-MIR-30E
hsa-mir-31 MIR-31
mmu-mir-31 MMU-MIR-31
hsa-mir-32 MIR-32
mmu-mir-32 MMU-MIR-32
hsa-mir-33a MIR-33A
hsa-mir-34c MIR-34C
mmu-mir-34c MMU-MIR-34C
hsa-mir-92a-1 MIR-92A-1
mmu-mir-92a-1 MMU-MIR-92A-1
hsa-mir-92a-2 MIR-92A-2
mmu-mir-92a-2 MMU-MIR-92A-2
mmu-mir-92b MMU-MIR-92B
mmu-mir-93 MMU-MIR-93
hsa-mir-95 MIR-95
hsa-mir-96 MIR-96
hsa-mir-28 MIR-28
mmu-mir-28 MMU-MIR-28
hsa-mir-29a MIR-29A
Precursor Name Catalog Number
hsa-mir-100 MIR-100
mmu-mir-100 MMU-MIR-100
mmu-mir-101a MMU-MIR-101A
mmu-mir-101b MMU-MIR-101B
hsa-mir-103-1 MIR-103-1
mmu-mir-103-1 MMU-MIR-103-1
hsa-mir-103-2 MIR-103-2
mmu-mir-105 MMU-MIR-105
hsa-mir-105-1 MIR-105-1
hsa-mir-105-2 MIR-105-2
hsa-mir-106a MIR-106A
mmu-mir-106a MMU-MIR-106A
hsa-mir-106b MIR-106B
mmu-mir-106b MMU-MIR-106B
hsa-mir-107 MIR-107
mmu-mir-107 MMU-MIR-107
mmu-mir-122 MMU-MIR-122
mmu-mir-124-1 MMU-MIR-124-1
hsa-mir-124-2 MIR-124-2
mmu-mir-124-2 MMU-MIR-124-2
mmu-mir-124-3 MMU-MIR-124-3
mmu-mir-125a MMU-MIR-125A
hsa-mir-125b-2 MIR-125B-2
mmu-mir-125b-2 MMU-MIR-125B-2
mmu-mir-126 MMU-MIR-126
mmu-mir-127 MMU-MIR-127
hsa-mir-128-1 MIR-128-1
mmu-mir-128-1 MMU-MIR-128-1
hsa-mir-128-2 MIR-128-2
mmu-mir-128-2 MMU-MIR-128-2
hsa-mir-129-1 MIR-129-1
mmu-mir-129-1 MMU-MIR-129-1
mmu-mir-96 MMU-MIR-96
hsa-mir-98 MIR-98
mmu-mir-98 MMU-MIR-98
Precursor Name Catalog Number
mmu-mir-130a MMU-MIR-130A
hsa-mir-130b MIR-130B
mmu-mir-130b MMU-MIR-130B
hsa-mir-132 MIR-132
mmu-mir-132 MMU-MIR-132
mmu-mir-133a-1 MMU-MIR-133A-1
mmu-mir-133a-2 MMU-MIR-133A-2
mmu-mir-133b MMU-MIR-133B
mmu-mir-134 MMU-MIR-134
hsa-mir-135a-1 MIR-135A-1
mmu-mir-135a-1 MMU-MIR-135A-1
hsa-mir-135a-2 MIR-135A-2
mmu-mir-135a-2 MMU-MIR-135A-2
hsa-mir-135b MIR-135B
mmu-mir-135b MMU-MIR-135B
hsa-mir-136 MIR-136
mmu-mir-136 MMU-MIR-136
hsa-mir-137 MIR-137
mmu-mir-137 MMU-MIR-137
hsa-mir-138-1 MIR-138-1
mmu-mir-138-1 MMU-MIR-138-1
hsa-mir-138-2 MIR-138-2
mmu-mir-138-2 MMU-MIR-138-2
hsa-mir-139 MIR-139
mmu-mir-139 MMU-MIR-139
hsa-mir-140 MIR-140
mmu-mir-140 MMU-MIR-140
mmu-mir-141 MMU-MIR-141
hsa-mir-142 MIR-142
mmu-mir-142 MMU-MIR-142
hsa-mir-143 MIR-143
mmu-mir-143 MMU-MIR-143
hsa-mir-129-2 MIR-129-2
mmu-mir-129-2 MMU-MIR-129-2
hsa-mir-130a MIR-130A
www.cellbiolabs.com [email protected]
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
76
Precursor Name Catalog Number
mmu-mir-146a MMU-MIR-146A
hsa-mir-146b MIR-146B
hsa-mir-147 MIR-147
hsa-mir-147b MIR-147B
hsa-mir-148a MIR-148A
mmu-mir-148a MMU-MIR-148A
mmu-mir-149 MMU-MIR-149
hsa-mir-150 MIR-150
mmu-mir-150 MMU-MIR-150
hsa-mir-151 MIR-151
mmu-mir-151 MMU-MIR-151
hsa-mir-152 MIR-152
mmu-mir-152 MMU-MIR-152
mmu-mir-153 MMU-MIR-153
hsa-mir-154 MIR-154
mmu-mir-154 MMU-MIR-154
mmu-mir-155 MMU-MIR-155
hsa-mir-181a-1 MIR-181A-1
mmu-mir-181a-1 MMU-MIR-181A-1
mmu-mir-181a-2 MMU-MIR-181A-2
mmu-mir-181b-1 MMU-MIR-181B-1
mmu-mir-181b-2 MMU-MIR-181B-2
mmu-mir-181c MMU-MIR-181C
hsa-mir-181d MIR-181D
mmu-mir-182 MMU-MIR-182
mmu-mir-183 MMU-MIR-183
mmu-mir-184 MMU-MIR-184
hsa-mir-185 MIR-185
mmu-mir-185 MMU-MIR-185
hsa-mir-186 MIR-186
hsa-mir-187 MIR-187
mmu-mir-187 MMU-MIR-187
mmu-mir-144 MMU-MIR-144
hsa-mir-145 MIR-145
mmu-mir-145 MMU-MIR-145
Precursor Name Catalog Number
mmu-mir-190b MMU-MIR-190B
mmu-mir-191 MMU-MIR-191
mmu-mir-192 MMU-MIR-192
mmu-mir-193 MMU-MIR-193
mmu-mir-193b MMU-MIR-193B
hsa-mir-194-1 MIR-194-1
mmu-mir-194-1 MMU-MIR-194-1
mmu-mir-194-2 MMU-MIR-194-2
hsa-mir-195 MIR-195
hsa-mir-196a-1 MIR-196A-1
mmu-mir-196a-1 MMU-MIR-196A-1
hsa-mir-196a-2 MIR-196A-2
mmu-mir-196a-2 MMU-MIR-196A-2
mmu-mir-196b MMU-MIR-196B
hsa-mir-197 MIR-197
hsa-mir-198 MIR-198
hsa-mir-199a-1 MIR-199A-1
mmu-mir-199a-1 MMU-MIR-199A-1
hsa-mir-199a-2 MIR-199A-2
hsa-mir-200a MIR-200A
mmu-mir-200a MMU-MIR-200A
mmu-mir-200b MMU-MIR-200B
hsa-mir-200c MIR-200C
mmu-mir-200c MMU-MIR-200C
mmu-mir-201 MMU-MIR-201
hsa-mir-202 MIR-202
mmu-mir-202 MMU-MIR-202
hsa-mir-204 MIR-204
mmu-mir-204 MMU-MIR-204
hsa-mir-205 MIR-205
mmu-mir-205 MMU-MIR-205
hsa-mir-206 MIR-206
mmu-mir-188 MMU-MIR-188
hsa-mir-190 MIR-190
mmu-mir-190 MMU-MIR-190
Precursor Name Catalog Number
mmu-mir-208b MMU-MIR-208B
mmu-mir-210 MMU-MIR-210
hsa-mir-211 MIR-211
mmu-mir-211 MMU-MIR-211
hsa-mir-212 MIR-212
mmu-mir-212 MMU-MIR-212
hsa-mir-214 MIR-214
mmu-mir-214 MMU-MIR-214
mmu-mir-215 MMU-MIR-215
hsa-mir-216a MIR-216A
mmu-mir-216a MMU-MIR-216A
mmu-mir-216b MMU-MIR-216B
mmu-mir-217 MMU-MIR-217
mmu-mir-218-1 MMU-MIR-218-1
mmu-mir-218-2 MMU-MIR-218-2
hsa-mir-219-1 MIR-219-1
mmu-mir-219-1 MMU-MIR-219-1
mmu-mir-219-2 MMU-MIR-219-2
hsa-mir-222 MIR-222
mmu-mir-222 MMU-MIR-222
mmu-mir-223 MMU-MIR-223
hsa-mir-224 MIR-224
mmu-mir-224 MMU-MIR-224
mmu-mir-290 MMU-MIR-290
mmu-mir-291a MMU-MIR-291A
mmu-mir-291b MMU-MIR-291B
mmu-mir-292 MMU-MIR-292
mmu-mir-293 MMU-MIR-293
mmu-mir-294 MMU-MIR-294
mmu-mir-295 MMU-MIR-295
mmu-mir-296 MMU-MIR-296
hsa-mir-297 MIR-297
mmu-mir-206 MMU-MIR-206
hsa-mir-208a MIR-208A
mmu-mir-208a MMU-MIR-208A
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
77
Precursor Name Catalog Number
mmu-mir-297a-5 MMU-MIR-297A-5
mmu-mir-297a-6 MMU-MIR-297A-6
mmu-mir-297b MMU-MIR-297B
mmu-mir-297c MMU-MIR-297C
hsa-mir-298 MIR-298
mmu-mir-298 MMU-MIR-298
mmu-mir-299 MMU-MIR-299
hsa-mir-301a MIR-301A
hsa-mir-301b MIR-301B
hsa-mir-302a MIR-302A
mmu-mir-302a MMU-MIR-302A
hsa-mir-302b MIR-302B
mmu-mir-302b MMU-MIR-302B
hsa-mir-302c MIR-302C
mmu-mir-302c MMU-MIR-302C
mmu-mir-302d MMU-MIR-302D
mmu-mir-320 MMU-MIR-320
mmu-mir-322 MMU-MIR-322
hsa-mir-323 MIR-323
mmu-mir-324 MMU-MIR-324
mmu-mir-325 MMU-MIR-325
mmu-mir-328 MMU-MIR-328
hsa-mir-329-1 MIR-329-1
mmu-mir-330 MMU-MIR-330
hsa-mir-331 MIR-331
mmu-mir-331 MMU-MIR-331
hsa-mir-335 MIR-335
mmu-mir-337 MMU-MIR-337
mmu-mir-338 MMU-MIR-338
mmu-mir-341 MMU-MIR-341
hsa-mir-342 MIR-342
mmu-mir-342 MMU-MIR-342
mmu-mir-297a-1 MMU-MIR-297A-1
mmu-mir-297a-3 MMU-MIR-297A-3
mmu-mir-297a-4 MMU-MIR-297A-4
Precursor Name Catalog Number
mmu-mir-345 MMU-MIR-345
mmu-mir-346 MMU-MIR-346
mmu-mir-351 MMU-MIR-351
mmu-mir-361 MMU-MIR-361
mmu-mir-363 MMU-MIR-363
mmu-mir-365-1 MMU-MIR-365-1
mmu-mir-365-2 MMU-MIR-365-2
mmu-mir-367 MMU-MIR-367
mmu-mir-370 MMU-MIR-370
mmu-mir-374 MMU-MIR-374
hsa-mir-374b MIR-374B
mmu-mir-375 MMU-MIR-375
mmu-mir-376a MMU-MIR-376A
mmu-mir-376c MMU-MIR-376C
mmu-mir-377 MMU-MIR-377
mmu-mir-378 MMU-MIR-378
mmu-mir-379 MMU-MIR-379
mmu-mir-380 MMU-MIR-380
mmu-mir-381 MMU-MIR-381
mmu-mir-382 MMU-MIR-382
mmu-mir-383 MMU-MIR-383
mmu-mir-384 MMU-MIR-384
mmu-mir-409 MMU-MIR-409
mmu-mir-410 MMU-MIR-410
mmu-mir-411 MMU-MIR-411
mmu-mir-412 MMU-MIR-412
mmu-mir-421 MMU-MIR-421
mmu-mir-423 MMU-MIR-423
mmu-mir-425 MMU-MIR-425
mmu-mir-429 MMU-MIR-429
mmu-mir-431 MMU-MIR-431
hsa-mir-433 MIR-433
mmu-mir-343 MMU-MIR-343
mmu-mir-344-1 MMU-MIR-344-1
mmu-mir-344-2 MMU-MIR-344-2
Precursor Name Catalog Number
mmu-mir-449a MMU-MIR-449A
mmu-mir-449b MMU-MIR-449B
mmu-mir-449c MMU-MIR-449C
mmu-mir-450a-2 MMU-MIR-450A-2
hsa-mir-450b MIR-450B
mmu-mir-451 MMU-MIR-451
mmu-mir-452 MMU-MIR-452
mmu-mir-455 MMU-MIR-455
mmu-mir-464 MMU-MIR-464
mmu-mir-465a MMU-MIR-465A
mmu-mir-465b-1 MMU-MIR-465B-1
mmu-mir-465b-2 MMU-MIR-465B-2
mmu-mir-465c-1 MMU-MIR-465C-1
mmu-mir-465c-2 MMU-MIR-465C-2
mmu-mir-466a MMU-MIR-466A
mmu-mir-466b-2 MMU-MIR-466B-2
mmu-mir-466b-3 MMU-MIR-466B-3
mmu-mir-466d MMU-MIR-466D
mmu-mir-466f-1 MMU-MIR-466F-1
mmu-mir-466f-4 MMU-MIR-466F-4
mmu-mir-466g MMU-MIR-466G
mmu-mir-466h MMU-MIR-466H
mmu-mir-466i MMU-MIR-466I
mmu-mir-466j MMU-MIR-466J
mmu-mir-466k MMU-MIR-466K
mmu-mir-466l MMU-MIR-466L
mmu-mir-467b MMU-MIR-467B
mmu-mir-467d MMU-MIR-467D
mmu-mir-467e MMU-MIR-467E
mmu-mir-467f MMU-MIR-467F
mmu-mir-467g MMU-MIR-467G
mmu-mir-469 MMU-MIR-469
mmu-mir-433 MMU-MIR-433
mmu-mir-434 MMU-MIR-434
mmu-mir-448 MMU-MIR-448
www.cellbiolabs.com [email protected]
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
78
Product Name Catalog Number
hsa-mir-603 MIR-603
hsa-mir-605 MIR-605
hsa-mir-606 MIR-606
hsa-mir-608 MIR-608
hsa-mir-609 MIR-609
hsa-mir-610 MIR-610
hsa-mir-613 MIR-613
hsa-mir-616 MIR-616
hsa-mir-619 MIR-619
hsa-mir-620 MIR-620
hsa-mir-628 MIR-628
hsa-mir-630 MIR-630
hsa-mir-633 MIR-633
hsa-mir-635 MIR-635
hsa-mir-636 MIR-636
hsa-mir-637 MIR-637
hsa-mir-641 MIR-641
hsa-mir-643 MIR-643
hsa-mir-645 MIR-645
hsa-mir-649 MIR-649
hsa-mir-651 MIR-651
hsa-mir-652 MIR-652
mmu-mir-652 MMU-MIR-652
hsa-mir-653 MIR-653
mmu-mir-653 MMU-MIR-653
hsa-mir-654 MIR-654
hsa-mir-658 MIR-658
hsa-mir-659 MIR-659
hsa-mir-665 MIR-665
mmu-mir-665 MMU-MIR-665
mmu-mir-666 MMU-MIR-666
mmu-mir-667 MMU-MIR-667
hsa-mir-600 MIR-600
hsa-mir-601 MIR-601
hsa-mir-602 MIR-602
Precursor Name Catalog Number
hsa-mir-543 MIR-543
mmu-mir-546 MMU-MIR-546
mmu-mir-547 MMU-MIR-547
hsa-mir-548a-1 MIR-548A-1
hsa-mir-548a-3 MIR-548A-3
hsa-mir-548b MIR-548B
hsa-mir-548c MIR-548C
hsa-mir-548d-1 MIR-548D-1
hsa-mir-548d-2 MIR-548D-2
hsa-mir-551b MIR-551B
hsa-mir-554 MIR-554
hsa-mir-555 MIR-555
hsa-mir-568 MIR-568
mmu-mir-568 MMU-MIR-568
hsa-mir-569 MIR-569
hsa-mir-577 MIR-577
hsa-mir-579 MIR-579
hsa-mir-580 MIR-580
hsa-mir-581 MIR-581
hsa-mir-582 MIR-582
mmu-mir-582 MMU-MIR-582
hsa-mir-584 MIR-584
hsa-mir-585 MIR-585
hsa-mir-591 MIR-591
hsa-mir-592 MIR-592
mmu-mir-592 MMU-MIR-592
hsa-mir-595 MIR-595
hsa-mir-596 MIR-596
hsa-mir-597 MIR-597
hsa-mir-598 MIR-598
mmu-mir-598 MMU-MIR-598
hsa-mir-599 MIR-599
mmu-mir-541 MMU-MIR-541
hsa-mir-542 MIR-542
mmu-mir-542 MMU-MIR-542
Precursor Name Catalog Number
mmu-mir-486 MMU-MIR-486
mmu-mir-487b MMU-MIR-487B
mmu-mir-488 MMU-MIR-488
mmu-mir-489 MMU-MIR-489
mmu-mir-490 MMU-MIR-490
hsa-mir-491 MIR-491
mmu-mir-491 MMU-MIR-491
mmu-mir-493 MMU-MIR-493
mmu-mir-494 MMU-MIR-494
mmu-mir-495 MMU-MIR-495
mmu-mir-496 MMU-MIR-496
mmu-mir-497 MMU-MIR-497
hsa-mir-499 MIR-499
mmu-mir-499 MMU-MIR-499
mmu-mir-501 MMU-MIR-501
hsa-mir-503 MIR-503
mmu-mir-503 MMU-MIR-503
hsa-mir-504 MIR-504
mmu-mir-504 MMU-MIR-504
hsa-mir-505 MIR-505
mmu-mir-505 MMU-MIR-505
hsa-mir-506 MIR-506
hsa-mir-508 MIR-508
hsa-mir-509-1 MIR-509-1
hsa-mir-514-1 MIR-514-1
hsa-mir-514-2 MIR-514-2
hsa-mir-520f MIR-520F
hsa-mir-525 MIR-525
hsa-mir-526a-2 MIR-526A-2
mmu-mir-539 MMU-MIR-539
mmu-mir-540 MMU-MIR-540
hsa-mir-541 MIR-541
mmu-mir-483 MMU-MIR-483
mmu-mir-484 MMU-MIR-484
mmu-mir-485 MMU-MIR-485
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
MICRORNA ANALYSIS miRNA Clone Collection
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
79
Product Name Catalog Number
mmu-mir-669c MMU-MIR-669C
mmu-mir-669d MMU-MIR-669D
mmu-mir-669e MMU-MIR-669E
mmu-mir-669g MMU-MIR-669G
mmu-mir-669h MMU-MIR-669H
mmu-mir-669j MMU-MIR-669J
mmu-mir-669k MMU-MIR-669K
mmu-mir-670 MMU-MIR-670
hsa-mir-671 MIR-671
mmu-mir-672 MMU-MIR-672
mmu-mir-674 MMU-MIR-674
mmu-mir-675 MMU-MIR-675
mmu-mir-676 MMU-MIR-676
mmu-mir-677 MMU-MIR-677
mmu-mir-679 MMU-MIR-679
mmu-mir-681 MMU-MIR-681
mmu-mir-682 MMU-MIR-682
mmu-mir-684-1 MMU-MIR-684-1
mmu-mir-684-2 MMU-MIR-684-2
mmu-mir-686 MMU-MIR-686
mmu-mir-688 MMU-MIR-688
mmu-mir-690 MMU-MIR-690
mmu-mir-694 MMU-MIR-694
mmu-mir-695 MMU-MIR-695
mmu-mir-697 MMU-MIR-697
mmu-mir-698 MMU-MIR-698
mmu-mir-699 MMU-MIR-699
mmu-mir-700 MMU-MIR-700
mmu-mir-701 MMU-MIR-701
mmu-mir-702 MMU-MIR-702
mmu-mir-703 MMU-MIR-703
mmu-mir-704 MMU-MIR-704
mmu-mir-668 MMU-MIR-668
mmu-mir-669a-3 MMU-MIR-669A-3
mmu-mir-669b MMU-MIR-669B
Product Name Catalog Number
mmu-mir-711 MMU-MIR-711
mmu-mir-713 MMU-MIR-713
mmu-mir-715 MMU-MIR-715
mmu-mir-717 MMU-MIR-717
mmu-mir-719 MMU-MIR-719
mmu-mir-720 MMU-MIR-720
mmu-mir-721 MMU-MIR-721
mmu-mir-741 MMU-MIR-741
mmu-mir-742 MMU-MIR-742
mmu-mir-743a MMU-MIR-743A
mmu-mir-743b MMU-MIR-743B
hsa-mir-744 MIR-744
mmu-mir-744 MMU-MIR-744
hsa-mir-758 MIR-758
mmu-mir-758 MMU-MIR-758
mmu-mir-759 MMU-MIR-759
mmu-mir-761 MMU-MIR-761
mmu-mir-763 MMU-MIR-763
mmu-mir-764 MMU-MIR-764
hsa-mir-766 MIR-766
hsa-mir-767 MIR-767
hsa-mir-770 MIR-770
mmu-mir-770 MMU-MIR-770
hsa-mir-802 MIR-802
mmu-mir-802 MMU-MIR-802
mmu-mir-804 MMU-MIR-804
mmu-mir-871 MMU-MIR-871
mmu-mir-872 MMU-MIR-872
mmu-mir-873 MMU-MIR-873
hsa-mir-874 MIR-874
mmu-mir-874 MMU-MIR-874
mmu-mir-875 MMU-MIR-875
mmu-mir-705 MMU-MIR-705
hsa-mir-708 MIR-708
mmu-mir-708 MMU-MIR-708
Product Name Catalog Number
mmu-mir-877 MMU-MIR-877
mmu-mir-878 MMU-MIR-878
mmu-mir-879 MMU-MIR-879
mmu-mir-880 MMU-MIR-880
mmu-mir-881 MMU-MIR-881
mmu-mir-883A MMU-MIR-883A
mmu-mir-883B MMU-MIR-883B
hsa-mir-885 MIR-885
hsa-mir-889 MIR-889
hsa-mir-891a MIR-891A
hsa-mir-892b MIR-892B
hsa-mir-920 MIR-920
hsa-mir-921 MIR-921
hsa-mir-922 MIR-922
hsa-mir-923 MIR-923
hsa-mir-924 MIR-924
hsa-mir-933 MIR-933
hsa-mir-934 MIR-934
hsa-mir-935 MIR-935
hsa-mir-936 MIR-936
hsa-mir-937 MIR-937
hsa-mir-938 MIR-938
hsa-mir-940 MIR-940
hsa-mir-941 MIR-941
hsa-mir-942 MIR-942
mmu-mir-1187 MMU-MIR-1187
mmu-mir-1188 MMU-MIR-1188
mmu-mir-1191 MMU-MIR-1191
mmu-mir-1192 MMU-MIR-1192
mmu-mir-1193 MMU-MIR-1193
mmu-mir-1195 MMU-MIR-1195
mmu-mir-1197 MMU-MIR-1197
hsa-mir-876 MIR-876
mmu-mir-876 MMU-MIR-876
hsa-mir-877 MIR-877
www.cellbiolabs.com [email protected]
80
Expression, Control, Reporter Vectors
miRNASelect™ Expression and Control Vectors
MICRORNA ANALYSIS
Product Name Size Catalog Number
miRNASelect™ pEGP-mir Cloning and Expression Vector 100 µL MIR-EXP-GP-C
miRNASelect™ pEP-mir Cloning and Expression Vector 100 µL MIR-EXP-C
Our miRNASelect™ Mammalian Expression Vectors provide an easy, efficient method to clone a miRNA precursor from any species. The desired miRNA sequence is cloned into a human ß-globin intron contained within the vector. Two vector formats are available: The pEP vector contains a puromycin selection
marker The pEGP vector contains a GFP-puromycin fu-
sion to allow selection by either marker Each expression vector is provided with a null (empty) control vector at no extra charge.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Recent Product Citations 1. Esposito, F. et al. (2012). Down-regulation of the miR-25 and miR-30d contributes to the development of anaplastic thyroid carcinoma
targeting the polycomb protein EZH2. J. Clin. Endocrinol. Metab. 97:E710-E718. (MIR-EXP-GP-C) 2. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog
pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-EXP-GP-C)
miRNASelect™ Null Control Vectors
Product Name Size Catalog Number
miRNASelect™ pEGP-mir Null Control Vector 100 µL MIR-NULL-GP
miRNASelect™ pEP-mir Null Control Vector 100 µL MIR-NULL
Our Null Control vectors have no cloned miRNA sequence. They are useful in conjunction with vectors from our miRNASelect™ Precursor Clone Collection.
Recent Product Citations 1. Dar, A. et al. (2013). The role of miR-18b in MDM2-p53 pathway signaling and melanoma progression. J. Natl. Cancer Inst. 105:433-442.
(MIR-NULL) 2. Larsen, M. et al. (2014). miRNA-130a regulates C/EBP-epsilon expression during granulopoiesis. Blood 123:1079-1089. (MIR-NULL-GP) 3. Yuan, L. et al. (2012). MicroRNA-212 displays tumor-promoting properties in on-small cell lung cancer cells and targets the hedgehog
pathway receptor PTCH1. Mol. Biol. Cell 23:1423-1434. (MIR-NULL-GP)
miRNASelect™ Precursor Clone Collection (Expression Vectors), cont.
Product Name Catalog Number
mmu-mir-1894 MMU-MIR-1894
mmu-mir-1895 MMU-MIR-1895
mmu-mir-1892 MMU-MIR-1892
mmu-mir-1224 MMU-MIR-1224
mmu-mir-1198 MMU-MIR-1198
Product Name Catalog Number
mmu-mir-1898 MMU-MIR-1898
mmu-mir-1899 MMU-MIR-1899
mmu-mir-1900 MMU-MIR-1900
mmu-mir-1897 MMU-MIR-1897
mmu-mir-1896 MMU-MIR-1896
Product Name Catalog Number
mmu-mir-1903 MMU-MIR-1903
mmu-mir-1904 MMU-MIR-1904
mmu-mir-1905 MMU-MIR-1905
mmu-mir-1907 MMU-MIR-1907
mmu-mir-1902 MMU-MIR-1902
MICRORNA ANALYSIS miRNA Viral Expression
81
RAPAd® miRNA Adenoviral Expression System
Product Name Size Catalog Number
1 Kit VPK-253 RAPAd® miRNA Adenoviral Expression System
RAPAd® Adenoviral Expression Systems produce recombinant adenovirus with substantial reduction in wild-type adenovirus, while doing so in a much shorter 2 week time frame. The RAPAd® miRNA Adenoviral Expression System is specifically designed to deliver miRNA sequences into your target cell.
Virtually No Wild-Type Virus: Backbone vector engineered to produce <300 wild-type plaques per 109 particles, compared with 104-106 WT plaques per 109 VP with most other methods
Faster Virus Production: Virus generated in 2 weeks compared to a few months with traditional methods
For more information on our complete selection of RAPAd® Adenovirus Ex-pression Systems for gene expression
studies, please see page 49.
www.cellbiolabs.com [email protected]
Recent Product Citation Li, P. et al. (2013). MicroRNA-638 is highly expressed in human vascular smooth muscle cells and inhibits PDGF-BB-induced cell proliferation and migration through targeting orphan nuclear re-ceptor NOR1. Cardiovasc. Res. 10.1093/cvr/cvt082. (VPK-253)
miRNA Retroviral Expression Vector
Product Name Size Catalog Number
10 µg RTV-017 pMXs-miR-GFP/Puro Retroviral Vector
Our pMXs-miR-GFP/Puro Retroviral Vector allows you to clone a miRNA sequence of interest for pack-aging into a recombinant retrovirus for delivery into a target cell.
For efficient packaging of your miRNA into an MMLV-based retrovirus, use one of our Platinum Retroviral Packaging Cell Lines found on page 64.
Recent Product Citation Mansour, M. et al. (2013). The TAL1 complex targets the FBXW7 tumor suppressor by activating miR-223 in human T cell acute lymphoblastic leukemia. J. Exp. Med. 210:1545-1557. (RTV-017)
Adenovirus Production using the RAPAd® Adenoviral Expression System.
Reporter System, Knockdown Enhancer MICRORNA ANALYSIS
82
RNAiBoost™ Reagent Kit for miRNA and siRNA Enhancement
Product Name Size Catalog Number
20 reactions RNAI-200
100 reactions RNAI-201 RNAiBoost™ Reagent Kit
RNA interference can occur in the presence of either siRNA or the mature form of miRNA. The RNABoost™ Reagent Kit increases the level of interference in the presence of siRNA. It also increases the rate of processing of pre-miRNA into mature miRNA.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
miRNASelect™ Functional Analysis Reporter System
Product Name Size Catalog Number
1 Kit MIR-GFP miRNASelect™ pMIR-GFP Reporter System
Assay Principle. If miRNA is present and binds to the 3’ UTR, translation of the GFP gene is repressed, resulting in loss of fluo-rescence.
The miRNASelect™ Functional Analysis Reporter System pro-vides a simple method for the evaluation of potential targets of miRNA. Binding of miRNA se-quences to their suspected tar-gets results in repression of trans-lation. In this system the miRNA target sequence, such as a 3’ UTR, is cloned into the provided pMIR-GFP vector in the multiple cloning sites immediately downstream of the GFP gene. If the miRNA is present and binds to the target sequence, translation is re-pressed and no green fluores-cence appears. If the miRNA does not bind the target se-quence, GFP translation occurs normally and green fluorescence may be seen.
OXIDATIVE STRESS / DAMAGE Oxidative Stress Overview
Measuring Oxidative Stress
84
Marker or
Type of Damage Sample Type
Cells Tissues Blood Urine Other
Protein Damage (p. 85-89)
Protein carbonyl content (PCC) X X X
3-Nitrotyrosine X X X
BPDE Protein Adduct X X X
Advanced Glycation End Products (AGE) X X X
Carboxyethyl Lynsine (CEL) X X X
Carboxymethyl Lynsine (CML) X X X
Methylglyoxal X X X
Advanced Oxidation Protein Products (AOPP) X X X
Lipid Peroxidation
(p. 90-93)
4-Hydroxynonenal (4-HNE) X X X
Malondialdehyde (MDA) X X X X
8-iso-Prostaglandin F2 (8-Isoprostane) X X X X
Oxidized Low Density Lipoprotein (OxLDL) X
DNA / RNA Damage and
Repair (p. 94-100)
8-hydroxyguanosine (8-OHG) X X X X Cerebrospinal Fluid
8-hydroxydeoxyguanosine (8-OHdG) X X X X
Abasic (AP) sites X X
Aldehyde DNA Damage (Etheno adducts) X X
BPDE DNA Adduct X X
Comet Assay X
Double-strand DNA breaks X
UV DNA Damage (CPD and 6-4PP) X
Reactive Oxygen Species
(p. 102-105)
Universal ROS X X X X
Hydrogen Peroxide X X X X
Nitric Oxide X X X X Saliva
Antioxidants & Antioxidant
Capacity (p. 106-110)
Superoxide Dismutase X X X X
Catalase X X X
Glutathione X X X X
Total Antioxidant Capacity (TAC) X X X X Food
Oxygen Radical Antioxidant Capacity (ORAC) X X X Food
Hydroxyl Radical Antioxidant Capacity (HORAC) X X X Food
Cellular Antioxidant Capacity (CAA) Antioxidant compounds
Oxidative stress may be measured using one of three primary methods:
Measure the reactive oxygen species (ROS) directly Measure the presence of antioxidants Measure the resulting damage to proteins, lipids, DNA or RNA (most reliable)
Use the following table to determine the best oxidative stress assays for your samples.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OXIDATIVE STRESS / DAMAGE Protein Damage
85
Assays and Reagents for Protein Damage
Cellular proteins are subject to damage in the presence of reactive oxygen species (ROS). The resulting protein damage may take the form of nitration or oxidation of various amino acid residues, or may result in formation of advanced glycation end products (AGE) or ad-vanced oxidation protein products (AOPP). We have developed unique assays to detect protein damage with higher sensitivity and more user-friendly protocols.
OxiSelect™ Nitrotyrosine Assay Kits and Antibodies
Product Name Detection Size Catalog Number
Nitrotyrosine ELISA Kit Colorimetric 96 Assays STA-305
5 x 96 Assays STA-305-5
Nitrotyrosine Immunoblot Kit Immunoblot/ECL 10 Blots STA-303
Goat Anti-Nitrotyrosine Polyclonal Antibody Immunoblot/ELISA 100 µg STA-003
Rabbit Anti-Nitrotyrosine Polyclonal Antibody Immunoblot/ELISA 100 µg STA-004
Protein Tyrosine Nitration Control (Nitrotyrosine-BSA) Immunoblot/ECL 10 µg STA-304
Formation of 3-Nitrotyrosine During Oxidative Stress.
Recent Product Citations 1. Toth, P. et al. (2014). Resveratrol treatment rescues neurovas-
cular coupling in aged mice: role of improved cerebromicrovas-cular endothelial function and downregulation of NADPH oxi-dase. Am. J. Physiol. Heart Circ. Physiol. 306:H299-H308. (STA-305)
2. Li, F.C. et al. (2013). Transition from oxidative stress to nitrosa-tive stress in rostral ventrolateral medulla underlies fatal intoxica-tion induced by organophosphate mevinphos. Toxicol. Sci. 135:202-217. (STA-305)
3. Medina, J.P. et al. (2013). Angiotensin receptor-mediated oxida-tive stress is associated with impaired cardiac redox signaling and mitochondrial function in insulin-resistant rats. Am. J. Physiol. Heart Circ. Physiol. 305:H599-H607. (STA-305)
4. Kong, X. et al. (2013). Pioglitazone enhances the blood pressure-lowering effect of losartan via synergistic attenuation of angio-tensin II-induced vasoconstriction. J. Renin Angiotensin Aldos-terone Syst. 10.1177/1170320313489061. (STA-305)
5. Lupachyk, S. et al. (2013). Endoplasmic reticulum stress plays a key role in the pathogenesis of diabetic peripheral neuropathy. Diabetes 62:944-952. (STA-305)
6. Cho, W.K. et al. (2013). IL-13 receptor 2-arginase 2 pathway mediates IL-13-induced pulmonary hypertension. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L112-L124. (STA-305)
7. Montez, P. et al. (2012). Angiotensin receptor blockade recovers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-305)
8. Tahrani, A.A. et al. (2012). Obstructive sleep apnea and diabetic neuropathy: a novel association in patients with type 2 diabetes. Am. J. Respir. Crit. Care Med. 186:434-441. (STA-305)
9. Suvakov, S. et al. (2012). Glutathione-S-transferase A1, M1, P1 and T1 null or low-activity genotypes are associated with en-hanced oxidative damage among haemodialysis patients. Nephrol. Dial. Transplant. 10.1093/ndt/gfs369. (STA-305)
10.Shevalye, H. et al. (2012). Metanx alleviates multiple manifesta-tions of peripheral neuropathy and increases intraepidermal nerve fiber density in Zucker diabetic fatty rats. Diabetes 61:2126-2133. (STA-305)
Our OxiSelect™ Nitrotyrosine Assay Kits provide a simple method to measure the formation of 3-nitrotyrosine in proteins. This assay is available in two formats: a 96-well competitive ELISA and an im-munoblot kit. The ELISA format can detect the pres-ence of 3-nitrotyrosine as low as 10 nM. Both kits can detect nitrotyrosine in protein from any species.
www.cellbiolabs.com [email protected]
86
OXIDATIVE STRESS / DAMAGE Protein Damage
OxiSelect™ Protein Carbonyl Assay Kits
Product Name Detection Size Catalog Number
OxiSelect™ Protein Carbonyl ELISA Kit 96 Assays STA-310
5 x 96 Assays STA-310-5
OxiSelect™ Protein Carbonyl Spectrophotometric Assay Spectrophotometric 40 Assays STA-315
OxiSelect™ Protein Carbonyl Immunoblot Kit Immunoblot/ECL 10 Blots STA-308
Colorimetric
Oxidized Protein Immunoblot Control (Carbonyl-BSA) Immunoblot/ECL 10 µg STA-309
OxiSelect™ Protein Carbonyl Fluorometric Assay Fluorometric 100 Assays STA-307
Protein Carbonyl ELISA Kit Sensitive: Detects samples as low as 10
µg/ml Greater Sample Retention: No concentration
or TCA precipitation steps that contribute to sample loss
Assay Principle for the OxiSelect™ Protein Oxidation Immunoblot Kit (STA-308).
Protein Carbonyl Immunoblot Kit No Molecular Weight Shift: DNPH derivatization
after immunoblotting allows direct comparison of oxidized and non-oxidized protein fingerprints
The most common products of protein oxidation in biological samples are the carbonyl derivatives of Pro, Arg, Lys and Thr residues. Such derivatives are chemically stable and serve as markers for oxidative stress in most types of reactive oxygen species. Our OxiSelect™ Protein Carbonyl Assay Kits provide rapid, efficient methods for detection of protein car-bonyls. Four assay formats are available: im-munoblot, ELISA, fluorometric and spectrophotomet-ric. All formats are suitable for use with purified pro-tein, plasma, serum, or cell lysate samples from any species.
Recent Product Citations 1. Cui, Z. et al. (2014). Identification of the immunoproteasome as a
novel regulator of skeletal muscle differentiation. Mol. Cell Biol. 34:96-109. (STA-308)
2. Pons, D. et al. (2012). Initial activation status of the antioxidant response determines sensitivity to carboplatin/paclitaxel treatment of ovarian cancer. Anticancer Res. 32:4723-4728. (STA-308)
3. Cristovao, A.C. et al. (2012). NADPH oxidase 1 mediates -synucleinopathy in Parkinsons’ Disease. J. Neurosci. 32:14465-14477. (STA-308)
4. Bitar, M. et al. (2012). Decline in DJ-1 and decreased nuclear translocation of Nrf2 in Fuchs endothelial corneal dystrophy. In-vest. Ophthalmol. Vis. Sci. 53:5806-5813. (STA-308)
5. Cannizzo, E. et al. (2012). Age-related oxidative stress compro-mises endosomal proteostasis. Cell Report 2(1):136-149. (STA-308)
6. Satapati, S. et al. (2012). Elevated TCA cycle function in the pa-thology of diet-induced hepatic insulin resistance and fatty liver. J. Lipid Res. 53:1080-1092. (STA-308)
7. Vulusevic, B. et al. (2014). Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZ-induced diabetic mice. Cardiovasc. Res. 101:306-316. (STA-310)
8. Dai, D.F. et al. (2013). Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides. Circ. Heart Fail. 6:1067-1076. (STA-310)
9. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A Biol. Sci. Med. Sci. 68:359-367. (STA-310)
10.Young, K. et al. (2013). Each to their own: skeletal muscles of different function use different biochemical strategies during aesti-vation at high temperature. J. Exp. Biol. 216:1012-1024. (STA-310)
11.Fishman , J. et al. (2012). Oxidative modification of the intestinal mucus layer is a critical but unrecognized component of trauma hemorrhagic shock-induced gut barrier failure. Am. J. Physiol. Gastrointest. Liver Physiol. 304:G57-G63. (STA-310)
12.Murakami, Y. et al. (2012). Receptor interacting protein kinase mediates necrotic cone but not rod cell death in a mouse model of inherited degeneration. PNAS 10.1073/pnas.1206937109. (STA-310)
13.Kang, K.A. et al. (2012). Baicalein inhibits oxidative stress-induced cellular damage via antioxidant effects. Toxic. And Ind. Health 28:412-421. (STA-310)
14.Gannon, A.M. et al. (2012). Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. Toxicol. Sci. 125:274-284. (STA-310)
15.Tang, H. et al. (2011). Intrinsic apoptosis in mechanically venti-lated human diaphragm: linkage to a novel Fox/FoxO1/Stat3-Bim axis. FASEB J. 25:2921-2936. (STA-310)
16.Robinson, C.K. et al. (2011). A major role for nonenzymatic anti-oxidant processes in the radioresistance of Halobacterium salina-rum. J. Bacteriol. 193:1653-1662. (STA-310)
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
87
OXIDATIVE STRESS / DAMAGE Protein Damage
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OxiSelect™ AOPP Assay Kit
Advanced oxidation protein products are toxins cre-ated during oxidative stress in patients with diabetes mellitus, atherosclerosis, renal complications, and HIV. Our OxiSelect™ AOPP Assay Kit provides a quick, easy method for assessing AOPP levels.
Product Name Detection Size Catalog Number
OxiSelect™ AOPP Assay Kit Colorimetric 200 Assays STA-318
50 µL STA-319 AOPP-Human Serum Albumin N/A
Fast: Obtain results in <30 minutes Sensitive: Detect concentrations as low as 5 µM
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Untreated Human Serum Albumin and AOPP-HSA Positive Control Tested with the OxiSelect™ AOPP Assay Kit.
OxiSelect™ BPDE Protein Adduct ELISA Kit
Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. They may also be found in some cooked foods. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reac-tions, benzo(a)pyrene is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both proteins and DNA. Our OxiSelect™ BPDE Protein Adduct ELISA Kit provides a convenient method to measure the modification of proteins by BPDE.
Sensitive: Detect concentrations as low as 60 ng/mL
Convenient: Quantify on a standard microplate reader
Versatile: Suitable for use with cell lysates, tis-sue homogenates, plasma or serum
Product Name Detection Size Catalog Number
OxiSelect™ BPDE Protein Adduct ELISA Kit Colorimetric 96 Assays STA-301
BPDE-BSA Standard Curve Generated Using the OxiSelect™ BPDE Protein Adduct ELISA Kit.
For information on our BPDE DNA Adduct ELISA Kit, please see page 99.
Recent Product Citations 1. Bloomer, R. et al. (2013). Safety profile of caffeine and 1,3-
dimethylamylamine supplementation in healthy men. Human and Exp. Toxicol. 10.1177/0960327113475680. (STA-318)
2. Park, S.H. et al. (2012). Effects of neutral pH and low-glucose degradation product-containing peritoneal dialysis fluid on sys-temic markers of inflammation and endothelial dysfunction: a randomized controlled 1-year follow-up study. Nephrol. Dial. Transp. 27:1191-1199. (STA-318)
3. Anderson, D. et al. (2010). Albumin-based microbubbles bind up-regulated scavenger receptors following vascular injury. J. Biol. Chem. 285:40645-40653. (STA-318)
OXIDATIVE STRESS / DAMAGE Protein Damage
88 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Advanced glycation end products (AGE) are formed during the Maillard reaction where reducing carbohy-drates react with lysine side chains and N-terminal amino groups of various macromolecules, particularly proteins. These AGE products can adversely affect the function of the affected proteins and play a role in atherosclerosis, diabetes, aging and renal disease. Our OxiSelect™ Advanced Glycation End Product Kits are designed for the rapid detection of AGE pro-tein adducts. We offer assays to study generic AGE formation or specific AGE strucutres including N-(Carboxyethyl) lysine (CEL), N-(Carboxymethyl) lysine (CML), and methylglyoxal (MG). All kits will detect AGE structures from protein of any species.
Sensitive: Detect levels as low as 1 µg/mL of AGE-protein adduct
Versatile: Compatible with cell lysates, plasma, serum, or purified proteins
Advanced Glycation End Products (AGE) Pathways.
OxiSelect™ Advanced Glycation End Product Kits & Antibodies
Product Name Detection Size Catalog Number
Colorimetric 96 Assays STA-817
5 x 96 Assays STA-817-5
Glycoaldehyde-AGE-BSA N/A 100 µg STA-348
OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit
OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit
Our OxiSelect™ Advanced Glycation End Product (AGE) Competitive ELISA Kit detects a variety of AGE structures including CML and pentosidine. It does not detect CEL or methylglyoxal (MG). Samples are added to a plate coated with an AGE-protein conjugate. AGE-protein adducts in the sample compete with the AGE-coated plate for antibody bind-ing. High AGE adduct content in a sample results in less binding of the antibody to the plate, producing a low signal.
Product Name Detection Size Catalog Number
OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit Colorimetric 96 Assays STA-813
CEL-BSA N/A 100 µg STA-302
OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit
Sensitive: Detect levels as low as 100 ng/mL of CEL-protein adduct
Versatile: Compatible with cell lysates, plasma, serum, or purified proteins
The OxiSelect™ N-(Carboxyethyl) Lysine (CEL) Competitive ELISA Kit detects CEL protein adducts in a variety of samples including cell lysates, blood sam-ples, and other protein sources.
OxiSelect™ AGE Competitive ELISA Kit Standard Curve.
OXIDATIVE STRESS / DAMAGE Protein Damage
89 www.cellbiolabs.com [email protected]
Product Name Detection Size Catalog Number
OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit 96 Assays STA-811
5 x 96 Assays STA-811-5
Mouse Anti-Methylglyoxal Monoclonal Antibody Immunoblot/
Immunohistochemistry 100 µg STA-011
MG-BSA N/A 100 µg STA-306
Colorimetric
OxiSelect™ N-(Carboxymethyl) Lysine (CML) Assays and Antibodies
Oxidized / Nitrated Proteins
Product Name Size Catalog Number
Copper (Cu++) Oxidized Human Low Density Lipoprotein (LDL) 100 µg STA-214
Malondialdehyde (MDA) Modified Human Albumin 100 µg STA-210
Malondialdehyde (MDA) Modified Human Apolipoprotein B-100 100 µg STA-211
Malondialdehyde (MDA) Modified Human Low Density Lipoprotein (LDL) 100 µg STA-212
Nitrated Human Low Density Lipoprotein (LDL) 100 µg STA-213
All proteins are provided at a concentration of 1.0 mg/mL.
Product Name Detection Size Catalog Number
Colorimetric 96 Assays STA-816
5 x 96 Assays STA-816-5
OxiSelect™ N-(Carboxymethyl) Lysine (CML) Immunoblot Kit Immunoblot 10 Blots STA-313
Goat Anti-N-CML Polyclonal Antibody Immunoblot/ELISA 100 µg STA-013
Rabbit Anti-N-CML Polyclonal Antibody Immunoblot/ELISA 100 µg STA-014
CML-BSA Control N/A 100 µg STA-314
OxiSelect™ N-(Carboxymethyl) Lysine (CML) Competitive ELISA Kit
OxiSelect™ Methylglyoxal (MG) Assays and Antibodies
Sensitive: Detect levels as low as 200 ng/mL of MG-protein adduct
Versatile: Compatible with cell lysates, plasma, serum, or purified proteins
The OxiSelect™ Methylglyoxal (MG) Competitive ELISA Kit detects MG protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources.
Sensitive: Detect levels as low as 3 ng/mL of CML-protein adduct with the ELISA kit
Versatile: Compatible with cell lysates, plasma, serum, or purified proteins
The OxiSelect™ N-(Carboxymethyl) Lysine (CML)Competitive ELISA Kit detects CML protein adducts in a variety of samples including cell lysates, blood samples, and other protein sources.
CML Protein Adduct Detected in Human Plasma with the OxiSelect™ CML Competitive ELISA Kit.
Assays and Reagents for Lipid Peroxidation
Lipid peroxidation is a well-defined mechanism of cellular damage in both animals and plants that occurs during aging and in some disease states. Our OxiSelect™ Lipid Peroxi-dation Assays allow you to quickly and easily quantify the most common markers and by-products of lipid peroxidation.
OxiSelect™ TBARS Assay Kit
The TBARS assay is a well-established method for screening and monitoring lipid peroxidation via the by-product malondialdehyde (MDA). MDA forms a 1:2 adduct with thiobarbituric acid. Our OxiSelect™ TBARS Assay Kit provides a more user-friendly protocol for quantitation of the MDA-TBA adduct compared to other commercial assays. This assay detects total MDA, both free and in pro-tein adducts, in a variety of samples including cell and tissue lysates, plasma, and urine.
Product Name Detection Size Catalog Number
Colorimetric or Fluorometric
200 Assays STA-330
5 x 200 Assays STA-330-5 OxiSelect™ TBARS Assay Kit (MDA Quantitation)
Fast: Obtain results in 30 minutes Sensitive: Smaller reaction volumes require less
sample; detect as little as 2 µM Convenient: 96-well format; no glass tubes are
required
90
OXIDATIVE STRESS / DAMAGE Lipid Peroxidation
Recent Product Citations 1. Chang, Q. et al. (2014). Cytochrome P450 2C epoxygenases
mediate photochemical stress-induced death of photoreceptors. J. Biol. Chem. 289:8337-8352.
2. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adap-tation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122.
3. Fu, L. et al. (2012). Ethyl pyruvate reduces ventilation-induced neutrophil infiltration and oxidative stress. J. Lipid Res. 53:1080-1092.
4. Brindeiro, C.M. et al. (2012). Tempol prevents altered K+ chan-nel regulation of afferent arteriolar tone in diabetic rat kidney. Hypertension 59:657-664.
5. Joshi, S.G. et al. (2011). Nonthermal dielectric-barrier discharge plasma-induced inactivation involves oxidative DNA damage and membrane lipid peroxidation in Escherichia coli. Antimicrob. Agents Chemother. 55:1053-1062.
6. Kasaikina, M.V. et al. (2011). Roles of the 15-kDa Selenoprotein (Sep15) in redox homeostasis and cataract development re-vealed by the analysis of Sep15 knockout mice. J. Biol. Chem. 286:33203-33212.
7. Fedeles, B.I. et al. (2011). Chemical genetics analysis of an aniline mustard anticancer reveals complex I of the electron transport chain as a target. J. Biol. Chem. 286:33910-33920.
8. Wojciechowski, P. et al. (2010). Resveratrol arrests and re-gresses the development of pressure overload- but not volume overload-induced cardiac hypertrophy in rats. J. Nutr. 140:962-968.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ MDA (Malondialdehyde) Assays and Antibodies
As a common by-product of lipid peroxidation, malondialdehyde (MDA) is a well-accepted marker of oxidative stress. Modification of proteins by MDA can cause structural and functional changes in oxidized proteins. We offer assays and antibodies to measure MDA in a variety of formats. Kits are available to measure total MDA as well as MDA protein adducts specifically.
Note: MDA is most reliably detected in fresh sam-ples, or in samples that have been frozen for a maxi-mum of 1-2 months. For samples stored for longer periods, consider testing other markers of lipid peroxi-dation such as 4-HNE or 8-isoprostane.
MDA-TBA Standard Curve Using a Standard Plate Reader.
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91
OXIDATIVE STRESS / DAMAGE Lipid Peroxidation
www.cellbiolabs.com [email protected]
OxiSelect™ MDA Adduct Assay Kits
Product Name Detection Size Catalog Number
Goat Anti-Malondialdehyde (MDA) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-031
Rabbit Anti-Malondialdehyde (MDA) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-032
Recent Product Citations 1. Montez, P. et al. (2012). Angiotensin receptor blockade recov-
ers hepatic UCP2 expression and aconitase and SDH activities and ameliorates hepatic oxidative damage in insulin resistant rats. Endocrinology 153:5845-5856. (STA-331)
2. Lazrak, A. et al. (2011). Regulation of alveolar epithelial Na+ channels by ERK1/2 in chlorine breathing mice. Am. J. Respir. Cell Mol. Biol. 10.1165/rcmb.2011-0309OC. (STA-331)
3. Zarogiannis, S.G. et al. (2011). Ascorbate and deferoxamine administration post chlorine exposure decrease mortality and lung injury in mice. Am. J. Respir. Cell Mol. Biol. 45:386-392. (STA-331)
4. Hall, J.A. et al. (2010). Absence of thyroid hormone activation during development underlies a permanent defect in adaptive thermogenesis. Endrocrinology 151:4573-4582. (STA-331)
5. Barabutis, N. et al. (2008). Antioxidant activity of growth hor-mone-releasing hormone antagonists in LNCaP human pros-tate cancer cell line. PNAS 105:20470-20475. (STA-331)
Sensitive: ELISA kit detects MDA protein ad-ducts as low as 6 pmol/mL
Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates
Immunoblot of MDA-BSA Control Using the OxiSelect™ MDA Immunoblot Kit. Immunoblot control was electroblotted onto a nitrocellulose membrane, followed by detection with the provided anti-MDA antibody.
Product Name Detection Size Catalog Number
OxiSelect™ MDA Immunoblot Kit Immunoblot 10 Blots STA-331
OxiSelect™ MDA Adduct Competitive ELISA Kit Colorimetric 96 Assays STA-832
5 x 96 Assays STA-832-5
MDA-BSA N/A 100 µg STA-333
OxiSelect™ MDA Polyclonal Antibodies
Our MDA Adduct Assay Kits provide simple methods to measuring these protein adducts in a variety of sample types. The MDA Adduct Competitive ELISA Kit is a sensitive method for the quantitation of MDA in proteins from cells, tissues, or blood. Samples are added to a malondialdehyde protein conjugate-coated plate. The MDA in the sample competes with the MDA on the plate for binding to the primary anti-MDA antibody. A high concentration of MDA in the sample results in little to no antibody binding to the plate, producing a low signal. Our MDA Immunoblot is a convenient method for qualitative measurement of MDA protein adducts.
OXIDATIVE STRESS / DAMAGE Lipid Peroxidation
92 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Standard Curve Generated with the OxiSelect™ HNE Adduct Competitive ELISA Kit.
OxiSelect™ HNE ELISA Kits and Antibodies
Product Name Detection Size Catalog Number
OxiSelect™ HNE Adduct Competitive ELISA Kit Colorimetric 96 Assays STA-838
5 x 96 Assays STA-838-5
Goat Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-034
Rabbit Anti-4-Hydroxynonenal (HNE) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-035
HNE-BSA N/A 100 µg STA-335
Sensitive: ELISA kit detects protein adducts as low as 2 µg/mL
Versatile: Suitable for use with serum, plasma, cell lysates or tissue homogenates
4-hydroxynonenal (4-HNE) is a well-known by-product of lipid peroxidation and is widely accepted as a stable marker for oxidative stress. HNE protein adducts are typically stable when frozen for up to 6 months or more. Our OxiSelect™ HNE Adduct Competitive ELISA Kit provides a simple, user-friendly way to assess HNE adduct formation on lysine, histidine and/or cysteine. Our polyclonal antibodies against HNE recognize HNE adducts to lysine, histidine, and/or cysteine and are suitable for use with Western Blot or ELISA ap-plications.
OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit (8-isoprostane)
Product Name Detection Size Catalog Number
OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit 96 Assays STA-337
5 x 96 Assays STA-337-5 Colorimetric
8-iso-Prostaglandin F2 is produced in membrane phospholipids and has been implicated in athero-genesis, rheumatoid arthritis and carcinogenesis. The OxiSelect™ 8-iso-Prostaglandin F2 ELISA Kit provides rapid, sensitive detection of 8-iso-PGF2as low as 50 pg/mL. The assay is suitable for quantita-tion of 8-isoprostane in a variety of sample types in-cluding cell and tissue lysates, plasma, serum, and urine.
Recent Product Citations 1. Dugas, T.R. et al. (2014). Hydrogen sulfide cytoprotective sig-
naling is endothelial nitric oxide synthase-nitric oxide dependent. PNAS 111:3182-3187.
2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Experimental Toxicol. 10.0177/0960327112472994.
3. Mollo, R. et al. (2012). Effect of -lipoic acid on platelet reactivity in type 1 diabetic patients. Diabetes Care 35:196-197.
4. Thompson, C.M. et al. (2012). Comparison of the effects of hexavalent chromium in the alimentary canal of F344 rats and B6C3F1 mice following exposure in drinking water: implications for carcinogenic modes of action. Toxicol. Sci. 125:79-90.
OXIDATIVE STRESS / DAMAGE Lipid Peroxidation
93 www.cellbiolabs.com [email protected]
OxiSelect™ Human Oxidized LDL ELISA Kits
LDL contains a hydrophobic core of various lipids surrounded by one molecule of Apolipoprotein B-100 (ApoB-100), which promotes solubility of the LDL in blood. LDL, often described as “bad” cholesterol, is even more dangerous when it becomes oxidized. Oxi-dized LDL (OxLDL) is more reactive with surrounding tissues and can collect within the inner lining of arter-ies. Our OxiSelect™ Human Oxidized LDL ELISA Kits are designed for the detection and quantitation of modi-fied LDL in human plasma or serum. Kits are avail-able to detect MDA-LDL, CML-LDL, or HNE-LDL in either the protein or lipid component of LDL. Our OxPL-LDL kit specifically detects oxidation in the phospholipid component of LDL.
Product Name Detection Size Catalog Number
OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation) Colorimetric 96 Assays STA-388
OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation) Colorimetric 96 Assays STA-389
OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation) Colorimetric 96 Assays STA-369
OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation) Colorimetric 96 Assays STA-358
Sensitive: Detect as little as 50 ng/mL of MDA-LDL, 150 ng/mL of CML-LDL, 150 ng/mL of HNE-LDL, or 100 ng/mL of OxPL-LDL
Quantitative: Compare unknown samples with provided copper oxidized LDL standard
Quantitation of MDA-LDL in Serum and Plasma Samples. Se-rum and plasma samples were treated with LDL Precipitation Solu-tion. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further diluting 1:160 in Assay Diluent according to the As-say Protocol. OxiSelect™ Human Oxidized LDL ELISA Assay Principle.
MDA is the most commonly found damage marker in oxidized LDL, but it can degrade in frozen samples after 1-2 months. CML and HNE, while less commonly found in OxLDL, may be more reliably detectable in samples that have been frozen for several months.
94
Assays for DNA & RNA Damage and Repair
DNA is arguably the most biologically significant target of oxidative and cellular stress. Continuous DNA damage has been implicated in age-related development of various can-cers. More recently, RNA damage has been described in conjunction with various neuro-logical diseases including Alzheimer’s and Parkinson’s diseases. We offer a wide range of assays to measure the most common types of DNA and RNA damage in cells.
OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation)
Among the various types of oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OHdG) is a ubiquitous marker of oxidative stress. 8-OHdG, one of the by-products of DNA oxidative damage, is physiologi-cally formed and enhanced by chemical carcino-gens. Our OxiSelect™ Oxidative DNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHdG in DNA samples. 8-OHdG is easily detect-able directly in serum and urine samples, after it is excised and secreted during the DNA repair proc-ess. It also may be detected in DNA extracted from cells or tissues of any species, following full DNA digestion into single bases.
Highly Sensitive: Detect as little as 100 pg/mL of 8-OHdG
Versatile: Suitable for use with urine, serum, and DNA extracted from cells or tissues
Product Name Detection Size Catalog Number
OxiSelect™ Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation) Colorimetric 96 Assays STA-320
5 x 96 Assays STA-320-5
Recent Product Citations 1. Moore, J. et al. (2013). Protection of corneal epithelial stem cells
prevents ultraviolet A damage during corneal collagen cross-linking treatment for keratoconus. Br. J. Ophthalmol. 10.1136/bjophthalmol-2013-303816.
2. Kalghatgi, S. et al. (2013). Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithe-lium of light-exposed mice. Sci. Transl. Med. 5:192ra85.
3. James, M.L. et al. (2013). VARA attenuates hyperoxia-induced impaired alveolar development and lung function in newborn mice. Am. J. Physiol. Lung Cell Mol. Physiol. 304:L803-L812.
4. Ravikumar, P. et al. (2013). Klotho protects lung epithelial cells against oxidative DNA damage. FASEB J. 27:722-723.
5. Singh, B. et al. (2013). MicroRNA-93 regulates NRF2 expression and is associated with breast carcinogenesis. Carcinogenesis 10.1093-carcin/bgt026.
6. Singh, B. et al. (2012). Superoxide dismutase 3 is induced by antioxidants, inhibits oxidative DNA damage and is associated with inhibition of estrogen-induced breast cancer. Carcinogene-sis 33:2601.2610.
7. Mercer, J.R. et al. (2012). The methyl xanthine caffeine inhibits DNA damage signaling and reactive species and reduces atherosclerosis in ApoE-/- mice. Arterioscler. Thromb. Vasc. Biol. 32:2461-2467.
8. Schutt, F. et al. (2012). Moderately reduced ATP levels promote oxidative stress and debilitate autophagic and phagocytic ca-pacities in human RPE cells. Invest. Ophthalmol. Vis. Sci. 53:5354-5361.
9. Tanabe, K. et al. (2012). Nicorandil as a novel therapy for ad-vanced diabetic nephrophathy in the eNOS-deficient mouse. Am. J. Physiol. Renal Physiol. 302:F1151-F1160.
10.Tzortzaki, E.G. et al. (2012). Oxidative DNA damage and so-matic mutations: a link to the molecular pathogenesis of chronic inflammatory airway diseases. Chest 141:1243-1250.
11.Xu, X. et al. (2012). Reactive oxygen species-triggered tro-phoblast apoptosis is initiated by endoplasmic reticulum stress via activation of caspase-12, CHOP, and the JNK pathway in Toxoplasma gondii infection in mice. Infect. Immun. 80:2121-2132.
12.Burnham, E. L. (2012). Protandim does not influence alveolar epithelial permeability or intrapulmonary oxidative stress in hu-man subjects with alcohol use disorders. Am. J. Physiol. Lung Cell Mol. Physiol. 302:L688-L699.
13.Pialoux, V. et al. (2011). Losartan abolishes oxidative stress induced by intermittent hypoxia in humans. J. Physiol. 589:5529-5537.
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OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
95
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
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OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation)
Similarly to 8-hydroxydeoxyguanosine (8-OHdG) forming during DNA oxidation, RNA can become oxidized resulting in 8-hydroxyguanosine (8-OHG). Oxidation of RNA has been implicated in a number of neurological diseases including Alzheimer’s and Parkinson’s diseases. Our OxiSelect™ Oxidative RNA Damage ELISA Kit provides a powerful method for rapid quantitation of 8-OHG in urine, serum or cerebrospinal fluid. It also may be used to detect 8-OHG in RNA extracted from cells or tissues of any species.
Highly Sensitive: Detect as little as 150 pg/mL of 8-OHG
Versatile: Suitable for use with urine, serum, cerebrospinal fluid, and DNA extracted from cells or tissues
Product Name Detection Size Catalog Number
OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation) Colorimetric 96 Assays STA-325
5 x 96 Assays STA-325-5
Recent Product Citations 1. Kannan, S. et al. (2012). Dendrimer-based postnatal therapy for
neuroinflammation and cerebral palsy in a rabbit model. Sci. Transl. Med. 4:130ra46.
2. Bazin, J. et al. (2011). Targeted mRNA oxidation regulates sun-flower seed dormancy alleviation during dry after-ripening. Plant Cell 23:2196-2208.
OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation)
Product Name Detection Size Catalog Number
OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine Quantitation) Colorimetric
96 Assays STA-825
5 x 96 Assays STA-825-5
Various reactive nitrogen species (RNS) including peroxynitrite and nitrogen oxides can form during pathophysiological conditions. These RNS can ni-trate guanine bases to form 8-nitroguanine in both DNA and RNA. Nitrosative damage to DNA and RNA is a significant contributor to the age-related development of major inflammation-related diseases as well as colon, breast, and prostate cancers. Our OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit provides a simple method for rapid quanti-tation of 8-nitroguanine in urine, serum or plasma samples. The assay measures total 8-nitroguanine from both DNA and RNA combined.
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Standard Curve Generated with the OxiSelect™ Nitrosative DNA/RNA Damage ELISA Kit (8-Nitroguanine).
Standard Curve Generated with the OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG).
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
96 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites)
Product Name Detection Size Catalog Number
OxiSelect™ Oxidative DNA Damage Quantitation Kit (AP Sites) Colorimetric 50 Assays STA-324
Highly Sensitive: Detect as few as 4-40 AP sites in 105 bp of DNA
Versatile: Suitable for use with genomic DNA from cells or tissues
Quantitative: Kit includes both oxidized and reduced DNA standards for absolute quantita-tion
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Standard Curve Generated with the OxiSelect™ Oxidative DNA Damage Quantitation Kit (STA-324).
OxiSelect™ DNA Double-Strand Break Assay
Product Name Detection Size Catalog Number
OxiSelect™ DNA Double-Strand Break Staining Kit Immuno-
fluorescence 100 Assays STA-321
Double-strand breaks (DSB) are among the most dangerous types of DNA damage within cells. An early cellular response is phosphorylation of the his-tone variant H2AX at the site of the DSB. This trig-gers a cascade of events and appears to play a role in recruitment of repair factors to the damaged sites. Our OxiSelect™ DNA Double-Strand Break Staining Kit provides an easy-to-use method for detecting DNA breaks. The kit utilizes simple immunofluores-cence staining of the phosphorylated histone H2AX.
Fast: See staining results in about 3 hours Positive Control: DNA Double-strand break
inducer included in kit
DNA Double-Strand Break Formation in A549 Cells. A549 cells were seeded at 50,000 cells/well overnight. Immunofluores-cence staining was then performed according to the assay pro-tocol. (A) Untreated cells. (B) Cells treated with 100 µM eto-poside for one hour.
Oxidative DNA Damage can manifest in the forma-tion of apurinic or apyrimidinic (AP) sites, also known as loss of bases. Spontaneous base loss, if unrepaired, can inhibit transcription and may be mutagenic. Our OxiSelect™ Oxidative DNA Damage Quantita-tion Kit provides a simple, user-friendly method for measuring AP sites in DNA. The assay uses an al-dehyde reactive probe (ARP) which specifically re-acts with an aldehyde group on the open ring of the AP site, followed by labeling with Biotin and subse-quent detection by Streptavidin-enzyme conjugate.
Recent Product Citations 1. Messaoudi, N. et al. (2013). Global stress response in a pro-
karyotic model of DJ-1-associated Parkinsonism. J. Bacteriol. 195:1167-1178.
2. Zaika, E. et al. (2011). P73 protein regulates DNA damage repair. FASEB J. 25:4406-4414.
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
97 www.cellbiolabs.com [email protected]
OxiSelect™ Comet Assays (Single Cell Gel Electrophoresis)
Product Name Detection Size Catalog Number
OxiSelect™ 3-Well Comet Assay Kit Light Microscopy
15 Wells STA-350
75 Wells STA-351
5 x 75 Wells STA-351-5
OxiSelect™ 3-Well Comet Assay Slides Light Microscopy
5 Slides STA-352
25 Slides STA-353
125 Slides STA-353-5
OxiSelect™ 96-Well Comet Assay Kit Light Microscopy 96 Wells STA-355
5 x 96 Wells STA-355-5
OxiSelect™ 96-Well Comet Assay Slides 1 Slide STA-356
5 Slides STA-356-5
OxiSelect™ Comet Assay Control Cells (includes positive and negative controls) N/A 1 Set STA-354
Light Microscopy
DNA damage can result from a variety of intracellu-lar and extracellular stimuli, and can manifest in a variety of mutations to the DNA including base modi-fications, missing bases and single-stranded or dou-ble-stranded breaks. Traditionally the comet assay, or single cell gel elec-trophoresis (SCGE), has been used as a well-published, high-level screening tool to measure DNA damage in single cells. Our OxiSelect™ Comet Assay Kits provide a quick, easy method to screen for DNA damage at a macro level. Our OxiSelect™ Comet Assay Slides have been specially treated for adhesion of low-melting agarose used in the assay. Damaged DNA moves farther in electrophoresis than intact DNA, causing a “tail” to form upon visualization under a fluorescence microscope.
Etoposide Treatment of Jurkat Cells. Jurkat cells were either untreated (left) or treated with etoposide (right) prior to perform-ing the OxiSelect™ Comet Assay.
Assay Principle for the OxiSelect™ Comet Assay Kit.
Recent Product Citations 1. Luo, Y. et al. (2013). SMC1-mediated intra-S-phase arrest
facilitates bocavirus DNA replication. J. Virol. 87:4017-4032. (STA-350, STA-351)
2. Li, Y.J. et al. (2012). Gold nanoparticles as a platform for creating a multivalent poly-SUMO chain inhibitor that also augments ionizing radiation. PNAS 109:4092-4097. (STA-350, STA-351)
3. Robin, T.P. et al. (2012). EWS/FLI1 regulates EYA3 in Ewing Sarcoma via modulation of miRNA-708, resulting in increased cell survival and chemoresistance. Mol. Cancer Res. 10:1098-1108. (STA-350, STA-351)
4. Choi, S.K. et al. (2012). Poly(ADP-ribose) polymerase 1 inhi-bition improves coronary arteriole function in type 2 diabetes mellitus. Hypertension 59:1060-1068. (STA-350, STA-351)
5. Tyagi, A. et al. (2011). Resveratrol selectively induces DNA damage, independent of Smad4 expression, in its efficacy against human head and neck squamous cell carcinoma. Clin. Cancer Res. 17:5402-5411. (STA-355)
98
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ UV-Induced DNA Damage Assays
Product Name Detection Size Catalog Number
OxiSelect™ UV-Induced DNA Damage ELISA Combo Kit (CPD/6-4PP) Colorimetric 96 Assays STA-322-C
OxiSelect™ UV-Induced DNA Damage ELISA Kit (CPD Quantitation) Colorimetric 96 Assays STA-322
5 x 96 Assays STA-322-5
Colorimetric 96 Assays STA-323
5 x 96 Assays STA-323-5 OxiSelect™ UV-Induced DNA Damage ELISA Kit (6-4PP Quantitation)
Product Name Detection Size Catalog Number
Colorimetric 96 Assays STA-326
5 x 96 Assays STA-326-5
OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (6-4PP) Colorimetric 96 Assays STA-328
OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kit (CPD)
OxiSelect™ UV-Induced DNA Damage ELISA Kits, for extracted DNA
Absorption of ultraviolet radiation can damage DNA by the formation of pyrimidine dimers. The two most common forms of pyrimidine dimers are cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimi-done photoproducts (6-4PP). Our OxiSelect™ UV-Induced DNA Damage Assays conveniently measure the formation of either CPD or 6-4PP in intact cells. Kits for each marker are avail-able in three formats: an ELISA for DNA extracted from cells or tissues, a Cell-Based ELISA, and a Cel-lular Immunostaining kit.
UV-Induced DNA Damage in HeLa Cells Treated with Ultravio-let Light for 30 Minutes and Visualized with the OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD).
UV-Induced DNA Damage in HeLa Cells as Detected with the OxiSelect™ Cellular UV-Induced DNA Damage ELISA (CPD).
Recent Product Citations 1. Zirkin, S. et al. (2013). The PIM-2 kinase is an essential com-
ponent of the ultraviolet damage response that acts upstream to E2F-1 and ATM. J. Biol. Chem. 288:21770-21789. (STA-322)
2. Burgess, H.M. et al. (2011). Nuclear relocalisation of cytoplas-mic poly(A)-binding proteins PABP1 and PABP4 in response to UV irradiation reveals mRNA-dependent export of meta-zoan PABPs. J. Cell Sci. 124:3344-3355. (STA-322)
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Product Name Detection Size Catalog Number
OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (CPD) Fluorescence Microscopy
96 Assays STA-327
OxiSelect™ Cellular UV-Induced DNA Damage Staining Kit (6-4PP) Fluorescence Microscopy
96 Assays STA-329
OxiSelect™ Cellular UV-Induced DNA Damage ELISA Kits, for intact cells
OxiSelect™ Cellular UV-Induced DNA Damage Staining Kits, for intact cells
99
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
www.cellbiolabs.com [email protected]
OxiSelect™ BPDE DNA Adduct ELISA Kit
Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, cigarette smoke, and automotive exhaust. One PAH, benzo(a)pyrene, was the first chemical carcinogen to be discovered. Through a series of enzymatic reac-tions, benzo(a)pyrene is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) which attacks both pro-teins and DNA. Our OxiSelect™ BPDE DNA Adduct ELISA Kit pro-vides a convenient method to measure BPDE ad-ducts in DNA extracted from cells or tissues.
Sensitive: Detect concentrations as low as 30 ng/mL
Convenient: Quantify on a standard microplate reader
Product Name Detection Size Catalog Number
OxiSelect™ BPDE DNA Adduct ELISA Kit Colorimetric 96 Assays STA-357
BPDE-DNA Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.
For information on our BPDE Protein Adduct ELISA Kit, please see page 87.
Product Name Detection Size Catalog Number
OxiSelect™ Aldehyde-Induced DNA Damage ELISA Combo Kit (Ethenoadenosine / Ethenocytidine Quantitation)
Colorimetric 96 Assays STA-820-C
OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenoadenosine Quantitation)
Colorimetric 96 Assays STA-820
OxiSelect™ Aldehyde-Induced DNA Damage ELISA Kit (Ethenocytidine Quantitation)
Colorimetric 96 Assays STA-821
OxiSelect™ Aldehyde-Induced DNA Damage Assays (Etheno Adducts)
Oxidation of phospholipids can lead to the formation of lipid hydroperoxides. These resulting short-lived hydrop-eroxides can either be converted to inert fatty acid alcohols, or can react with metals to form aldehydes such as malondialdehyde (MDA), 4-hydroxynonenal (HNE), acrolein, and crotonaldehyde. These aldehydes (which can also be formed through exposure to carcinogenic substances such as urethane or vinyl chloride) can damage DNA resulting in the formation of various etheno adducts, including 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine. The presence of these bases can lead to base pair substitution mutations.
Our OxiSelect™ Aldehyde-Induced DNA Damage Assays conveniently measure the formation of either 1,N6-ethenodeoxyadenosine (ethenoadenosine) or 3,N4-ethenodeoxycytidine (ethenocytidine) in DNA extracted from cells or tissues. In addition, we offer a convenient combination kit that can measure both etheno bases in separate wells of the same plate.
Etheno Base Structures that Form Adducts with DNA During Oxidative Stress.
Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epox-ide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/kfu003.
100 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OXIDATIVE STRESS / DAMAGE DNA / RNA Damage & Repair
Checkpoint Kinase Activity Assays
Product Name Detection Size Catalog Number
Checkpoint Kinase Activity Immunoblot Kit Immunoblot 20 Assays STA-413
Colorimetric 96 Assays STA-414
5 x 96 Assays STA-414-5 96-Well Checkpoint Kinase Activity Assay Kit
Checkpoint kinases, specifically CHK1 and CHK2, are activated in response to DNA damage to subsequently phosphorylate Cdc25C prior to mitosis, which prompts cell cycle arrest. Mutation of these checkpoint kinases can ultimately lead to decreased DNA repair. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phospho-specific antibody. Checkpoint Kinase Activitiy Assays are available in two formats: a Western blot assay and a 96-well plate-based activity assay.
Product Name Detection Size Catalog Number
Global DNA Methylation ELISA Kit (5’-methyl-2’-deoxycytidine Quantitation)
Colorimetric 96 Assays STA-380
5 x 96 Assays STA-380-5
Global DNA Methylation ELISA Kit
Our Global DNA Methylation and Hydroxymethyla-tion Assays provide a convenient, accurate way to quantify 5’-methyl-2’-deoxycytidine (5MedCyd) and 5-hydroxymethylcytosine respectively. Unknown samples are compared with a standard provided with each kit.
Sensitive: Detect as little as 15 nM of 5MedCyd Versatile: Suitable for use with any isolated
DNA as well as urine samples Convenient: Quantify on a standard microplate
reader
Methylated DNA Standard Curve Generated with the Global DNA Methylation ELISA Kit.
5MedCyd Levels in Human Urine Sample as Measured with the Global DNA Methylation ELISA Kit.
DNA methylation is an epigenetic change shown to be associated with nearly every biological process. In mammalian cells, DNA methylation is found predominantly at CpG dinucleotides; however, in certain cases such as embryonic stem cells it may also be found in non-CpG contexts. Due to the important role of DNA me-thylation in maintaining genomic stability, deregulation of DNA methylation is associated with various diseases including cancer.
101 www.cellbiolabs.com [email protected]
OXIDATIVE STRESS / DAMAGE Hypoxia Assays
HIF-1 Alpha DNA Binding Activity Assay Kit
Detection Specificity of HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM deferoxamine mesylate (DFO) for 4 hours at 37ºC. Nuclear extracts were prepared using the Nuclear/Cytosolic Fractionation Kit (#AKR-171). 100 pmol of non-biotinylated wild type or mutated HRE double stranded com-petitor oligos were added to the Complete DNA Binding Buffer just prior to inclusion in the assay.
Product Name Detection Size Catalog Number
HIF-1 Alpha DNA Binding Activity Assay Kit Colorimetric 96 Assays CBA-282
Cell hypoxia, or low oxygen condition, is a normal physiological response to certain body stressors such as high altitudes, but it also can be a symptom of pathological conditions. In some cases hypoxia may contribute to the inducement of oxidative stress. In response to hypoxic conditions, the hypoxia-inducible factor 1 transcriptional activator complex (HIF-1) plays a role in activating several hypoxia-responsive genes such as erythropoietin and VEGF. During hy-poxia, the alpha subunit of HIF-1 accumulates and translocates from the cytosol to the nucleus, where it dimerizes with the beta subunit and becomes tran-scriptionally active. It then binds transcriptional coacti-vators to induce gene expression. The HIF-1 Alpha DNA Binding Activity Assay Kit is an ELISA-based assay to detect activated HIF-1. Active HIF-1 complex is captured on a double-stranded oligo containing a hypoxic response element (HRE) that is attached to the plate. Detection is then performed with a primary antibody followed by an HRP-conjugated secondary antibody. The assay will detect HIF-1 complexes from human, mouse or rat samples.
Product Name Detection Size Catalog Number
HIF-1 Alpha Sandwich ELISA Kit Colorimetric 96 Assays CBA-280
HIF-1 Alpha Cell Based ELISA Kit Chemiluminescent 96 Assays CBA-281
HIF-1 Alpha ELISA Kits
Detection of Nuclear HIF-1 Alpha. HeLa cells were incubated in the presence or absence of 0.2 mM DFO for 4 hours at 37ºC. Nu-clear extracts were prepared using the Nuclear/Cytosolic Fractiona-tion Kit. HIF-1 Alpha levels were measured in untreated (blue bars) and treated (red bars) extracts according to the Assay Protocol.
Our HIF-1 Alpha ELISA Kits provide a convenient method for detection and quantitation of human, mouse, or rat HIF-1 Alpha in cells or tissues. Two ELISA kit formats are available: The HIF-1 Alpha Sandwich ELISA Kit detects HIF
-1 Alpha in any protein sample including tissue homogenates, whole cell lysates, or nuclear ex-tracts. Samples are added to an anti-HIF-1 Alpha antibody coated plate. Quantitation of unknown samples is performed by comparison of the OD values to those of a known standard.
The HIF-1 Alpha Cell Based ELISA Kit allows the detection of HIF-1 Alpha levels in intact cells. Cells are seeded in a tissue culture treated plate suitable for reading in a 96-well plate-based lumi-nometer. Cells are fixed and permeabilized to allow detection with the anti-HIF-1 antibody. De-tection is performed by chemiluminescence.
OXIDATIVE STRESS / DAMAGE ROS Assays
Reactive Oxygen Species Assays
Reactive oxygen species (ROS) such as superoxide and hydrogen peroxide are continually produced during metabolic processes. Excess ROS can lead to cellular injury in the form of damaged DNA, lipids and proteins. We offer assays for quantitation of various reactive oxy-gen species, in both in vitro and intracellular formats.
OxiSelect™ Intracellular ROS Assay Kit
The OxiSelect™ Intracellular ROS Assay Kit measures the activity of hydroxyl, peroxyl, and other reactive oxy-gen species. The assay uses the cell-permeable fluoro-genic probe DCFH-DA, which diffuses into cells and is deacetylated into the non-fluorescent DCFH. In the presence of ROS, the DCFH is oxidized into highly fluorescent DCF. Fluorescence is quantified on a fluorometric plate reader.
Sensitive: Detect concentrations as little as 10 pM Fast: Entire protocol takes about one hour
Product Name Detection Size Catalog Number
Fluorometric 96 Assays STA-342
5 x 96 Assays STA-342-5 OxiSelect™ Intracellular ROS Assay Kit
Assay Principle for the OxiSelect™ Intracellular ROS Assay.
Recent Product Citations 1. Koontz, J. et al. (2014). Competition through dimerization be-
tween antiapoptotic and proapoptotic HS-1-associated protein X-1 (Hax-1). J. Biol. Chem. 289:3468-3477.
2. Kim, E.Y. et al. (2013). NOX2 interacts with podocyte TRPC6 channels and contributes to their activation by diacylglycerol: essential role of podocin in formation of this complex. Am. J. Physiol. Cell Physiol. 305:C960-C971.
3. Abe, Y. et al. (2013). TGF-ß1 stimulates mitochondrial oxidative phosphorylation and generation of reactive oxygen species in cultured mouse podocytes, mediated in part by the mTOR path-way. Am. J. Phyisol. Renal Physiol. 305:F1477-F1490.
4. He, Q. et al. (2013). Tafazzin knockdown interrupts cell cycle progression in cultured neonatal ventricular fibroblasts. Am. J. Physiol. Heart Circ. Physiol. 305:H1332-H1343.
5. Kokkinaki, M. et al. (2013). Klotho regulates retinal pigment epithelial functions and protects against oxidative stress. J. Neu-rosci. 33: 16346-16359.
6. Hagan, C. et al. (2013). A common docking domain in progester-one receptor-B links DUSP6 and CK2 signaling to proliferative transcriptional programs in breast cancer cells. Nucleic Acids Res. 10.1093/nar/gkt706.
7. Teng, H. et al. (2013). Oxygen-sensitive mitochondrial accumu-lation of cystathione ß-synthase mediated by lon protease. PNAS 110:12679-12684.
8. Wang, H. et al. (2012). p53-induced gene 3 mediates cell death induced by glutathione peroxidase 3. J. Biol. Chem. 287:16890-16902.
9. Druz, A. et al. (2012). Glucose depletion activates MMU-mir-466h-5p expression through oxidative stress and inhibition of histone deacetylation. Nucleic Acids Res. 10.1093/nar/gks452.
10.Montalvo-Ortiz, B.L. et al. (2012). Characterization of EHOP-016, novel small molecule inhibitor of Rac GTPase. J. Biol. Chem. 287:13228-13238.
102 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OXIDATIVE STRESS / DAMAGE ROS Assays
103 www.cellbiolabs.com [email protected]
Product Name Detection Size Catalog Number
Fluorometric 96 Assays STA-347
5 x 96 Assays STA-347-5 OxiSelect™ In Vitro ROS/RNS Assay Kit
OxiSelect™ In Vitro ROS/RNS Assay Kit
Free radicals and related reactive oxygen species (ROS) and reactive nitrogen species (RNS) can ap-pear in the body both inside and outside the cell. Until recently it has been difficult to detect ROS and RNS outside of intact cells. The OxiSelect™ In Vitro ROS/RNS Assay Kit allows you to measure ROS and RNS formation in various body fluids including urine, serum and plasma. It is also useful for testing cell lysates, tissue homoge-nates, and cell culture supernatants. The assay universally measures reactive oxygen and reactive nitrogen species that may include hydrogen peroxide, nitric oxide, peroxynitrite, peroxyl radicals, and others. The assay principle is similar to our Intra-cellular ROS Assay (previous page), except that the chemistry is modified to allow detection of ROS out-side the cell. Fluorescence is quantified on a fluoro-metric plate reader.
Sensitive: Detect concentrations as little as 10 pM for DCF or 40 nM for hydrogen peroxide
Fast: Entire protocol takes about one hour Versatile: Suitable for a wide variety of sample
types including urine, serum, plasma, cell lysates, tissue homogenates and cell culture supernatants
Recent Product Citations 1. Liu, X. et al. (2013). Epoxyeicosatrienoic acids prevent cisplatin-
induced renal apoptosis through a p38 mitogen-activated protein kinase-regulated mitochondrial pathway. Mol. Pharmacol. 84:925-934.
2. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adap-tation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122.
3. Xiao, D. et al. (2013). Estrogen normalizes perinatal nicotine-induced hypertensive responses in adult female rat offspring. Hypertension 61:1246-1252.
4. Wang, W. et al. (2012). Mono-(2-ethylhexyl) phthalate induces oxidative stress and inhibits growth of mouse ovarian antral follicles. Biol. Reprod. 87:152.
5. Ju, D. J. et al. (2012). Ethyl pyruvate ameliorates albuminuria and glomerular injury in the animal model of diabetic nephropa-thy. Am. J. Phyisol. Renal Physiol. 302:F606-F613.
6. Momi, S. et al. (2012). Nitric oxide enhances the anti-inflammatory and anti-atherogenic activity of atorvastatin in a mouse model of accelerated atherosclerosis. Cardiovasc. Res. 10.1093/cvr/cvs100.
7. Patterson, A.J. et al. (2011). Hypoxia-derived oxidative stress mediates epigenetic repression of PKC epsilon gene in foetal rat hearts. Cardiovasc. Res. 10.1093/cvr/cvr322.
8. Rathnasamy, G. et al. (2011). Iron and iron regulatory proteins in amoeboid microglial cells are linked to oligodendrocyte death in hypoxic neonatal rat periventricular white matter through produc-tion of proinflammatory cytokines and reactive oxygen/nitrogen species. J. Neurosci. 31:17982-17995.
Assay Principle for the OxiSelect™ In Vitro ROS/RNS Assay.
OXIDATIVE STRESS / DAMAGE ROS Assays
104 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ Hydrogen Peroxide Assay, Colorimetric
Hydrogen peroxide is one of the most prevalent and most stable of the various reactive oxygen species. The half-life of hydrogen peroxide is significantly longer than that of most ROS, making it easier to detect in many sample types.
Standard Curve Generated with the OxiSelect™ Hydrogen Peroxide/Peroxidase Assay (Fluorometric).
Product Name Detection Size Catalog Number
OxiSelect™ Hydrogen Peroxide Assay Kit Colorimetric 500 Assays STA-343
Product Name Detection Size Catalog Number
OxiSelect™ Hydrogen Peroxide/Peroxidase Assay Kit Fluorometric 500 Assays STA-344
*For the testing of hydrogen peroxide in cells and tissues, please see our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit below.
OxiSelect™ Hydrogen Peroxide / Peroxidase Assay, Fluorometric
Our OxiSelect™ Hydrogen Peroxide Assay Kit pro-vides a simple method for quantitation of hydrogen peroxide. This colorimetric assay measures the oxi-dation of ferrous (Fe2+) ions to ferric (Fe3+) ions in the presence of peroxides. The ferric ions form a com-plex with a provided dye which may be read on a standard microplate reader. The assay may be run with either aqueous phase or lipid phase samples.
Recent Product Citations 1. Kim, E.Y. et al. (2012). Sustained activation of N-methyl-D-
aspartate receptors in podocytes leads to oxidative stress, mobi-lization of transient receptor potential canonical 6 channels, nuclear factor of activated T cells activation, and apoptotic cell death. Mol. Pharmacol. 82:728-737.
2. Kim, E.Y. et al. (2012). Insulin increases surface expression of TRPC6 channels in podocytes: role of NADPH oxidases and reactive oxygen species. Am. J. Phyisol. Renal Physiol. 302:F298-F307.
Sensitive: Detect as little as 50 nM Fast: Easy 30 minute incubation Versatile: Measure either hydrogen peroxide or
peroxidase in plasma, serum, urine, cell culture supernatants, cell lysates and tissue homogenates
Our OxiSelect™ Hydrogen Peroxide / Peroxidase Assay Kit provides a convenient plate-based method for quantitation of hydrogen peroxide or peroxidases in a wide variety of sample types. This fluorometric assay uses a fluorogenic probe which is converted from a non-fluorescent to a fluo-rescent state in the presence of peroxides and is catalyzed by peroxidases. The kit includes both a hydrogen peroxide standard and a peroxidase standard for quantitative results with either target.
Sensitive: Detect as little as 1 µM Fast: Easy 30-90 minute incubation, depending on
sample type Versatile: Suitable for plasma, serum, urine, and
cell culture supernatants*
OXIDATIVE STRESS / DAMAGE ROS Assays
105 www.cellbiolabs.com [email protected]
Direct detection: Probe binds directly to nitric ox-ide, not to by-products such as nitrate and nitrite
Sensitive: Detect as little as 3 nM Versatile: Read results as endpoint or time course
(kinetic) in a fluorescence plate reader, or visualize under a fluorescence microscope
OxiSelect™ Intracellular Nitric Oxide Assay Kit
OxiSelect™ In Vitro Nitric Oxide Assay Kits
Product Name Detection Size Catalog Number
Fluorometric 96 Assays STA-800
5 x 96 Assays STA-800-5 OxiSelect™ Intracellular Nitric Oxide (NO) Assay Kit
Product Name Detection Size Catalog Number
OxiSelect™ In Vitro Nitric Oxide (Nitrite / Nitrate) Assay Kit
Colorimetric 100 Assays STA-802
5 x 100 Assays STA-802-5
100 Assays STA-801
5 x 100 Assays STA-801-5 Fluorometric
Nitric oxide (NO) is a progenitor of various reactive nitrogen species (RNS) in conjunction with superox-ide anions via nitric oxide synthase (NOS). It plays a role in vascular diseases, diabetes, atherosclerosis, inflammatory diseases and cancer. Because of its short half-life, nitric oxide is often difficult to detect directly. The OxiSelect™ Intracellular Nitric Oxide Assay Kit allows direct detection of NO in intact cells. A cell-permeable fluorogenic probe is added to cells; upon treatment to induce oxidative stress, nitric oxide that is generated within the cell binds to the probe pro-ducing a bright fluorescent signal. Results may be visualized under a fluorescence microscope or quantified in a 96-well fluorescence plate reader. Induction of NOS in RAW 264.7 Cells. Cells were seeded in a
96-well plate at 100,000 cells/well. Cells were uninduced (left) or induced with 50 ng/mL LPS and 10 ng/mL IFN(right) for 20 hours at 37ºC.
Nitric oxide (NO) is difficult to detect directly in vitro due to its short half-life. It is therefore common to measure nitric oxide formation by detection of its final oxidized products, nitrate and nitrite. The OxiSelect™ In Vitro Nitric Oxide Assay Kits pro-vide a convenient plate-based method for the quanti-tation of nitrate and nitrite in a variety of sample types. First, nitrate is reduced to nitrite. Then total nitrite is measured by the addition of a Griess Re-agent (for colorimetric detection) or a fluorometric probe (for fluorescence detection). Results are then quantified in a 96-well plate reader. OxiSelect™ In Vitro Nitric Oxide Assay Kits are suit-able for use with serum, plasma, urine, saliva, cell lysates, and culture media.
Assay Principle for the OxiSelect™ In Vitro Nitric Oxide (Nitite / Nitrate) Assay, Fluorometric Format.
106
OXIDATIVE STRESS / DAMAGE Antioxidant Assays
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Antioxidant Assays
ROS generation is normally counterbalanced by the action of antioxidant enzymes and other redox molecules. We offer two types of assays for antioxidant quantitation:
OxiSelect™ Catalase Activity Assay Kits
Product Name Detection Size Catalog Number
Colorimetric 96 Assays STA-341
Fluorometric 96 Assays STA-339 OxiSelect™ Catalase Activity Assay Kit
Catalase is a ubiquitous enzyme that destroys hydro-gen peroxides formed during oxidative stress. Since hydrogen peroxides have a longer half-life than most free radicals and can make up a large portion of all reactive oxygen species, the ability to remove hydro-gen peroxides can be extremely important at combat-ing oxidative stress. Our OxiSelect™ Catalase Activity Assay Kits provide a quick, user-friendly protocol to monitor catalase activity from a variety of sample types. Kits are avail-able with either colorimetric or fluorometric detection.
Sensitive: Detect as little as 1.25 units/mL (colorimetric) or 50 mU/mL (fluorometric)
Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with whole blood,
plasma, serum, cell lysates or tissue homogenates High Throughput: 96-well format
Assays to quantify the presence or activity of antioxidant molecules Assays to determine the antioxidant capacity of biomolecules
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Assay Principle for the OxiSelect™ Catalase Acitivity Assays. Catalase present in samples converts hydrogen peroxide into water and oxygen (Reaction 1). Any remaining hydrogen peroxide that is not converted reacts with a colorimetric or fluorometric probe in the presence of horseradish peroxidase (Reaction 2) to produce a color or fluorescence which is measured in a plate reader.
Standard Curve Generated with the OxiSelect™ Catalase Activity Assay Kit (Colorimetric).
Recent Product Citation Costantini, D. et al. (2013). Loss of integration is associated with reduced resistance to oxidative stress. J. Exp. Biol. 216:2213-2220. (STA-341)
OXIDATIVE STRESS / DAMAGE Antioxidant Assays
107 www.cellbiolabs.com [email protected]
OxiSelect™ Superoxide Dismutase Activity Assay
Superoxide dismutase (SOD), which catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen, is one of the most important antioxidant enzymes.
Sensitive: Detect as little as 0.6 units/mL Fast: Obtain results in about 2 hours Versatile: Suitable for use with urine, serum, cells
or tissue samples
Product Name Detection Size Catalog Number
OxiSelect™ Superoxide Dismutase Activity Assay Colorimetric 100 Assays STA-340
Standard Curve Using the OxiSelect™ Superoxide Dismutase Activity Assay.
The OxiSelect™ Superoxide Dismutase Activity As-say uses a xanthine/xanthine oxidase (XOD) system to generate superoxide anions and a chromagen to produce a water-soluble dye upon reduction by the superoxide anions.
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OxiSelect™ Superoxide Dismutase Activity Assay Principle. Superoxide anions generated by a Xanthine/Xanthine Oxidase system are detected with the provided chromagen. SOD reduces superoxide concentrations, so higher SOD concentrations result in a decreased signal.
Recent Product Citations 1. Wysocki, J. et al. (2014). ACE2 deficiency increases NADPH-
mediated oxidative stress in teh kidney. PHY2 2:e00264. 2. Yin, J. et al. (2014). Development of an antioxidant system after
early weaning in piglets. J. Anim. Sci. 92:612-619. 3. Gong, E.J. et al. (2013). Low-dose-rate radiation exposure leads
to testicular damage with decreases in DNMT1 and HDAC1 in the murine testis. J. Radiat. Res. 10.1093/jrr/rrt090.
4. Song, J. et al. (2013). Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adap-tation. J. Gerontol. A Biol. Sci. Med. Sci. 10.1093/gerona/glt122.
5. Paneni, F. et al. (2013). Deletion of the activated protein-1 tran-scription factor JunD induces oxidative stress and accelerates age-related endothelial dysfunction. Circulation 127:1229-1240.
6. Connell, B.J. et al. (2012). UPEI-100, a conjugate of lipoic acid and apocynin, mediates neuroprotection in a rat model of ische-mia/reperfusion. Am. J. Physiol. Regulatory Integrative Comp. Physiol. 302:R866-R895.
7. Zhang, Z. et al. (2012). TRPM2 Ca2+ channel regulates energy balance and glucose metabolism. Am. J. Phyisol. Endocrin. Metab. 302:E807-E816.
OXIDATIVE STRESS / DAMAGE Antioxidant Assays
108 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit
Glutathione is an intracellular tripeptide thiol that pro-tects cells from free radicals by acting as an antioxi-dant. Glutathione exists within cells in both reduced (GSH) and oxidized (GSSG) forms; it is involved in the breakdown of peroxides and also helps maintain exogenous antioxidants such as vitamins C and E. The OxiSelect™ Total Glutathione Assay Kit is a quantitative assay for measuring total combined GSH and GSSG content in a variety of sample types. Oxi-dized glutathione is enzymatically reduced, followed by colorimetric detection in a microplate reader.
Sensitive: Detect as little as 8 nM total glutathione Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with saliva, urine,
serum, plasma, and cell or tissue lysates
Product Name Detection Size Catalog Number
OxiSelect™ Total Glutathione (GSSG/GSH) Assay Kit Colorimetric 100 Assays STA-312
OxiSelect™ Glutathione Reductase Assay Kit
Product Name Detection Size Catalog Number
OxiSelect™ Glutathione Reductase Assay Kit Colorimetric 100 Assays STA-812
Assay Principle for the OxiSelect™ Total Glutathione Assay Kit. In the presence of NADPH, glutathione reductase (provided) con-verts all glutathione into reduced form (GSH). The reduced glutathione then reacts with the provided chromogen to yield a color detect-able at 405 nm.
Standard Curve Generated with the OxiSelect™ Glutathione Reductase Assay Kit. Various concentrations of glutathione reductase standard were tested according to the Assay Protocol. OD values were read at 1 minute increments at 405 nm.
The OxiSelect™ Glutathione Reductase Assay Kit is a quantitative assay for measuring the activity levels of glutathione reductase in a variety of sample types. The assay principle is similar to that of our Total Glu-tathione Assay Kit above, except that endogenous levels of glutathione reductase drive the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH).
Sensitive: Detect activity levels as low as 0.6 mU/mL
Fast: Obtain results in less than 30 minutes Versatile: Suitable for use with erythrocytes,
plasma, cell lysates, or tissue extracts
Recent Product Citations 1. Mani, S. et al. (2013). Decreased endogenous production of
hydrogen sulfide accclerates atherosclerosis. Circulation 127:2523-2534.
2. Karakus, E. et al. (2013). Agomelatine: an antidepressant with new potent hepatoprotective effects on paracetamol-induced liver damage in rats. Human and Exp. Toxicol. 10.01177/0960327112472994.
OXIDATIVE STRESS / DAMAGE Antioxidant Assays
109 www.cellbiolabs.com [email protected]
OxiSelect™ Cellular Antioxidant Assay Kit, for in vivo Evaluation of Exogenous Antioxidants
Product Name Detection Size Catalog Number
OxiSelect™ Cellular Antioxidant Activity Assay Kit Fluorometric 192 Assays STA-349
Measuring the effects of antioxidant compounds in an in vitro assay may not accurately reflect their efficacy because such assays do not account for physiologi-cal conditions such as pH, temperature, uptake, me-tabolism, or the bioavailability or efficacy of an anti-oxidant compound. The OxiSelect™ Cellular Antioxidant Activity Assay Kit provides a mechanism to test exogenous antioxi-dants in a cell-based environment, delivering a more accurate measurement of the compound’s true physiological efficacy. A cell-permeable fluorometric dye is added to intact cells; when free radicals are generated, they bind to the dye producing a bright fluorescent signal. When the exogenous antioxidant is added, it eliminates the free radicals resulting in decreased fluorescence.
Physiological: Measures the efficacy of an antioxi-dant compound in a cellular environment
More Accurate: Compared to in vitro assays that do not take into account pH, temperature, or other relevant intracellular conditions
Sensitive: Detects as little as 10 µM Quercetin, a bioflavonoid which serves as the assay’s standard
Fast: Obtain results in about one hour
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Cellular Antioxidant Activity of Quercetin in HeLa Cells. 60,000 HeLa cells were seeded and cultured in a 96-well plate until confluent. Cells were then pretreated with DCFH-DA and Quercetin for 60 minutes at 37ºC. Free Radical Initiator was then added to the cells to begin the assay. Fluorescence readings were taken every 5 minutes for one hour at 37ºC.
Assay Principle for the OxiSelect™ Cellular Antioxidant Activ-ity Assay Kit. An exogenous antioxidant compound is added to cells along with DCFH-DA dye. Upon entry into the cell, the DCFH-DA is cleaved to DCFH which can bind reactive oxygen species (ROS) generated within the cell by the addition of a free radical initiator. Binding of DCFH to ROS yields DCF which produces a bright fluorescence. The presence of the exogenous antioxidant compound reduces the ROS available to the DCFH dye, yielding a lower fluorescent signal.
OXIDATIVE STRESS / DAMAGE Antioxidant Assays
110 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
OxiSelect™ ORAC and HORAC Activity Assay Kits
The ORAC (Oxygen Radical Antioxidant Capacity) and HORAC (Hydroxyl Radical Antioxidant Capac-ity) assays measure the antioxidant capacity of bio-molecules against peroxyl radicals and hydroxyl radicals, respectively. The assays are suitable for plasma, cell fractions, and tissue lysates, as well as solid and aqueous nutrition samples.
Assay Principle for the OxiSelect™ Oxygen Radical Antioxidant Capacity (ORAC) Assay.
Product Name Detection Size Catalog Number
Fluorometric 192 Assays STA-345
5 x 192 Assays STA-345-5
OxiSelect™ HORAC Activity Assay Kit Fluorometric 192 Assays STA-346
5 x 192 Assays STA-346-5
OxiSelect™ ORAC Activity Assay Kit
Recent Product Citations 1. Ungvari, Z. et al. (2013). Testing predictions of the oxidative
stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J. Gerontol. A. Biol. Sci. Med. Sci. 68:359-367.
2. Bailey-Downs, L.C. et al. (2011). Liver-specific knockdown of IGF-1 decreases vascular oxidative stress resistance by impair-ing the Nrf2-dependent antioxidant response: a novel model of vascular aging. J. Gerontol. A. Biol. Sci. Med. Sci. 10.1093/gerona/glr164.
Product Name Detection Size Catalog Number
OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit Colorimetric 200 Assays STA-360
Recent Product Citations 1. Stark, M. et al. (2013). Differential effects of docosahexaenoic
acid on preterm and term placental pro-oxidant/antioxidant bal-ance. Reproduction 146:243-251.
2. Bakalova, R. et al. (2013). Tissue redox activity as a hallmark of carcinogenesis: from early to terminal stages of cancer. Clin. Cancer Res. 19:2503-2517.
3. Wang, Y. et al. (2013). Therapeutic effect of MG-132 on diabetic cardiomyopathy is associated with its suppression of protea-somal activities: roles of Nrf2 and NF-kB. Am. J. Physiol. Heart Circ. Physiol. 304:H567-H578.
OxiSelect™ Total Antioxidant Capacity Assay Principle.
OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit
The OxiSelect™ Total Antioxidant Capacity (TAC) Assay Kit measures the total antioxidant capacity of biomolecules from a variety of sample types via a Single Electron Transfer (SET) mechanism. The as-say works with a variety of antioxidants and is suit-able for testing plasma, serum, urine, cell lysates, tissue homogenates and food extracts.
112
CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein
Small GTPase Activation Assay Principle.
Safe: Non-radioactive assay format Visual Check: Agarose beads can be easily seen Fast Results: 1 hour plus electrophoresis/blotting
Small GTP-binding proteins (GTPases) regulate a variety of cell signaling pathways and are therefore involved in a wide range of cell functions, processes, and morphology. The most studied small GTPases include Ras, Rac, Rho and Cdc42. We offer a variety of tools to enable the study of these small GTPase family members:
In addition, we offer sensitive assays to detect cyclic AMP and cyclic GMP, both of which are important regulators in the G-Protein signaling cascade.
Small GTPase / G-Protein Signaling
Small GTPase Activation Assays Small GTPase Activation ELISA Kits Small GTPase Assay Beads Active Rac-GEF Assay
Small GTPase Expression Vectors Small GTPase Premade Adenoviruses Small GTPase Retroviral Constructs Small GTPase Recombinant Proteins
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Small GTPase Activation Assays
Our Small GTPase Activation Assays use visible aga-rose beads to selectively pull down the active form of the target of interest. The precipitated GTPase is then detected by Western blot using a target specific anti-body included in the kit. If you are studying more than one small GTPase tar-get, consider one of our Small GTPase Activation Assay Combo Kits. These combo kits allow you to measure the following targets at a savings compared to buying separate kits for each target: Rac1 and Cdc42 RhoA, Rac1 and Cdc42
Visible Agarose Beads Provided in the Small GTPase Activation Assays. Beads are easy to visual-ize, making it easier to avoid poten-tial loss during washes and aspira-tions.
113
Small GTPase/G-Protein
Small GTPase Activation Assays (continued)
Product Name Detection Size Catalog Number
Cdc42 Activation Assay Immunoblot/ECL 20 Assays STA-402
Rac1 Activation Assay Immunoblot/ECL 20 Assays STA-401-1
Ral Activation Assay Immunoblot/ECL 20 Assays STA-408
Ran Activation Assay Immunoblot/ECL 20 Assays STA-409
Pan-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400
RhoA Activation Assay Immunoblot/ECL 20 Assays STA-403-A
Rac1/Cdc42 Activation Assay Combo Kit Immunoblot/ECL 20 Assays/Target STA-404
RhoA/Rac1/Cdc42 Activation Assay Combo Kit Immunoblot/ECL 10 Assays/Target STA-405
Rac2 Activation Assay Immunoblot/ECL 20 Assays STA-401-2
RhoC Activation Assay Immunoblot/ECL 20 Assays STA-403-C
Arf6 Activation Assay Immunoblot/ECL 20 Assays STA-407-6
Arf1 Activation Assay Immunoblot/ECL 20 Assays STA-407-1
Rap1 Activation Assay Immunoblot/ECL 20 Assays STA-406-1
RhoB Activation Assay Immunoblot/ECL 20 Assays STA-403-B
Rap2 Activation Assay Immunoblot/ECL 20 Assays STA-406-2
H-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-H
K-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-K
N-Ras Activation Assay Immunoblot/ECL 20 Assays STA-400-N
Recent Product Citations 1. Ohashi, K. et al. (2012). Lung cancers with acquired resistance to EGFR inhibitors occasionally harbor BRAF gene mutations but lack mu-
tations in KRAS, NRAS or MEK1. PNAS 109:E2127-E2133. (STA-400) 2. Geryk-Hall, M. et al. (2010). Driven to death: inhibition of farnesylation increases Ras activity in osteosarcoma and promotes growth arrest
and cell death. Mol. Cancer Ther. 9:1111-1119. (STA-400) 3. Camalier, C.E. et al. (2010). Elevated phosphate activates N-ras and promotes cell transformation and skin tumorigenesis. Cancer Prev.
Res. 3:359-370. (STA-400) 4. Tsukamoto, Y. et al. (2013). A novel heart failure mice model of hypertensive heart disease by angiotensin II infusion, nephrectomy, and
salt loading. Am. J. Physiol. Heart Circ. Physiol. 305:H1658-H1667. (STA-401-1) 5. Petzold, T. et al. (2013). ß1 integrin-mediated signals are required for platelet granule secretion and hemostasis in mouse. Blood 122:2723-
2731. (STA-401-1) 6. Cristante, E. et al. (2013). Identification of an essential endogenous regulator of blood-brain barrier integrity, and its pathological and thera-
peutic implications. PNAS 110:832-841. (STA-401-1) 7. Yanagashita, T. et al. (2014). Actin-binding protein, espin: a novel metastatic regulator for melanoma. Mol. Cancer Res. 12:440-446. (STA-
401-1, STA-403-A) 8. He, S. et al. (2013). SRGAP1 is a candidate gene for papillary thyroid carcinoma susceptibility. J. Clin. Endocrinol. Metab. 98:E973-E980.
(STA-402) 9. Fiuza, M. et al. (2013). GluN3A expression restricts spine maturation via inhibition of GIT1/Rac1 signaling. PNAS 110:20807-20812. (STA-
403-A) 10. Basu, M. et al. (2012). Wnt/ß-catenin pathway is regulated by PITX2 homeodomain protein and thus contributes to the proliferation of hu-
man ovarian adenocarcinoma cell SKOV-3. J. Biol. Chem. 288:4355-4367. (STA-403-A) 11. Holmes, K.M. et al. (2012). Insulin-like growth factor-binding protein 2-driven glioma progression is prevented by blocking a clinically signifi-
cant integrin, integrin-linked kinase, and NF-kB network. PNAS 109:2168-2173. (STA-404) 12. Wang, J. et al. (2013). DEK depletion negatively regulates Rho/ROCK/MLC pathway in non-small cell lung cancer. J. Histochem. & Cyto-
chem. 10.1369/00221155413488120. (STA-405) 13. Quint, P. et al. (2013). Sphingosine 1-phosphate (S1P) receptors 1 and 2 coordinately induce mesenchymal cell migration through S1P
activation of complementary kinase pathways. J. Biol. Chem. 288:5398-5406. (STA-405) 14. Baranwai, S. et al. (2011). Molecular characterization of the tumor-suppressive function of nischarin in breast cancer. J. Natl. Cancer Inst.
10.1093/jnci/djr350. (STA-405) 15. Grossman, A. et al. (2013). The small GTPase ARF6 stimulates ß-catenin transcriptional activity during WNT5A-mediated melanoma inva-
sion and metastasis. Sci Signal 6:ra14. (STA-407-6)
www.cellbiolabs.com [email protected]
CELL SIGNALING & PROTEIN BIOLOGY
114
Product Name Detection Size Catalog Number
Active Rac-GEF Assay Kit (Tiam1) Immunoblot/ECL 20 Assays STA-422
Active Rac-GEF Assay Kit (Tiam1)
Guanine nucleotide exchange factors (GEFs) acti-vate small GTPases by catalyzing the exchange of GDP for GTP. Our Active Rac-GEF Assay Kit (Tiam1) uses the agarose bead technology of our Small GTPase Activation Assays (previous page). Agarose beads pull down the active form of Rac-GEFs from en-dogenous lysates or purified samples. The specific GEF known as Tiam1 is then specifically detected with a polyclonal antibody.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein
Recent Product Citation Oubaha, M. et al. (2012). Formation of a PKC/ß-catenin complex in endothelial cells promotes angiopoietin-1-induced collective direc-tional migration and angiogenic sprouting. Blood 120:3371-3381.
Tiam1 Activation Assay. Left: 293 cells were transfected with active Tiam1. Active Tiam1 in lysate was pulled down with Rac1 G15A agarose beads. Right: Active Tiam-1 in 2 mg of MDA-231 cell lysate was pulled down with Rac1 G15A agarose beads and probed with anti-Tiam1 anti-body.
96-Well Ras Activation ELISA Kits
Product Name Detection Size Catalog Number
Colorimetric 96 Assays STA-440
Chemiluminescent 96 Assays STA-441 96-Well Ras Activation ELISA Kit
Our 96-Well Ras Activation Assays use the Raf1 Rho binding domain (Raf1 RBD) to selectively pull down the active form of Ras from purified or endogenous samples. The captured GTP-Ras is then detected by a pan-Ras antibody and HRP-conjugated secondary antibody. Detection is by either colorimetric or chemiluminescent plate reader.
EGF Stimulation and Active Ras Detection with the 96-Well Ras Activation ELISA Kit. HeLa cells were serum starved for 18 hours before EGF stimulation of 50 ng/mL for 2 minutes. Lysates were then prepared according to the assay protocol.
Pan-Ras Antibody Specificity. Anti-pan-Ras antibody reactivity with H-Ras, K-Ras and N-Ras human isoforms by dot blot.
115
Product Name Target Size Catalog Number
Cdc42 G15A Agarose Beads Cdc42-GEF 800 µg STA-433
Rac1 G15A Agarose Beads Rac1-GEF 800 µg STA-432
RhoA G17A Agarose Beads RhoA-GEF 400 µg STA-431
GEF Agarose Assay Beads
Recent Product Citations 1. Yuen, H. et al. (2013). RanGTPase: a candidate for Myc-
mediated cancer progression. J. Natl. Cancer Inst. 105:475-488. (STA-410)
2. Moniz, S. et al. (2007). Protein kinase WNK2 inhibits cell prolif-eration by negatively modulating the activation of MEK1/ERK1/2. Oncogene 26(41):6071-6081. (STA-410)
3. Pothula, S. et al. (2013). Regulation of Cdc42 expression and signaling is critical for promoting corneal epithelial wound heal-ing. Invest. Ophthalmol. Vis. Sci. 54:5343-5352. (STA-411)
4. Zhang, Q-G. et al. (2009). Estrogen attenuates ischemic oxida-tive damage via an estrogen receptor alpha-mediated inhibition of NADPH oxidase activation. J. Neurosci. 29:13823-13836. (STA-411)
5. Levy-Adam, F. et al. (2008). Heparanase facilitates cell adhe-sion and spreading by clustering of cell surface heparan sulfate proteoglycans. PLoS ONE 3(6):e2319. (STA-411, STA-412)
6. Sabbatini, M. E. et al. (2010). CCK activates RhoA and Rac1 differentially through G-alpha-13 and G-alpha-q in mouse pan-creatic acini. Am. J. Physiol. Cell Physiol. 298:C592-C605. (STA-411, STA-412)
7. Sabbatini, M. et al. (2008). Rap1 activation plays a regulatory role in pancreatic amylase secretion. J. Biol. Chem. 283:23884-23894. (STA-412)
www.cellbiolabs.com [email protected]
Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY
Small GTPase Agarose Assay Beads
Our agarose beads are useful for selectively pulling down only the active form of small GTPases. The beads are colored for easily visualization. These are the same beads used in our Small GTPase Ac-tivation Assays (p. 108-109).
Visible Agarose Beads. Beads are easy to visualize, making it easier to avoid potential loss during washes and aspirations.
Recent Product Citations 1. Ngok, S. et al. (2013). Phosphorylation-mediated 14-3-3 protein
binding regulates the function of the Rho-specific guanine nu-cleotide exchange factor (RhoGEF) Syx. J. Biol. Chem. 288:6640-6650. (STA-431)
2. Colacios, C. et al. (2011). The p.Arg63Trp polymorphism con-trols Vav1 functions and Fox3p regulatory T cell development. J. Exp. Med. 208:2183-2191. (STA-432)
Product Name Target Size Catalog Number
GGA3 PBD Agarose Beads Arf 400 µg STA-419
PAK1 PBD Agarose Beads Cdc42, Rac 400 µg STA-411
Raf1 RBD Agarose Beads Ras 400 µg STA-410
RalBP1 PBD Agarose Beads Ral 400 µg STA-420
RalGDS RBD Agarose Beads Rap 400 µg STA-418
RanBP1 Agarose Beads Ran 400 µg STA-421
Rhotekin RBD Agarose Beads Rho 400 µg STA-412
Our GEF agarose beads are useful for selectively pulling down only the active form of guanine nu-cleotide exchange factors (GEF). The beads are colored for easily visualization.
CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein
116 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Product Name Vectors Size Catalog Number
Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-455
GFP-Cdc42 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-451
Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 100 µL STA-454
GFP-Rac1 Expression Vector Set Wild Type, T17N (Dom. Neg.), Q61L (Active) 3 x 100 µL STA-450
H-Ras Expression Vector Set Wild Type, T17N (Dom. Neg.), G12V (Active) 3 x 100 µL STA-457
RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), G14V (Active) 3 x 100 µL STA-456
GFP-RhoA Expression Vector Set Wild Type, T19N (Dom. Neg.), Q63L (Active) 3 x 100 µL STA-452
Product Name Size Catalog Number
3 x 100 µL STA-460 Exoenzyme C3 Expression Vector
Small GTPase Expression Vector Sets
Our Small GTPase Expression Vectors are ideal tools for the study of the most commonly researched small GTPase targets. Each set contains 3 mammal-ian expression vectors: Wild type Dominant negative Constitutively active Vectors are available with or without a GFP reporter gene. All vectors are supplied as bacterial glycerol stocks.
Exoenzyme C3 (Rho Inhibitor) Expression Vector
Product Name Vectors Size Catalog Number
Active Rac1 Expression Vector Set Q61L, Q61L/F37A, Q61L/Y40C 3 x 100 µL STA-458
Active H-Ras Expression Vector Set V12, V12/S35, V12/C40 3 x 100 µL STA-459
Active Small GTPase Expression Vector Sets
Our Active Small GTPase Expression Vectors are similar to the expression vectors above, except that they are provided as sets of 3 different active mutants for a single small GTPase target. All vectors are supplied as bacterial glycerol stocks.
This vector is supplied as bacterial glycerol stock.
117 www.cellbiolabs.com [email protected]
Recent Product Citation Zhao, B. et al. (2012). TNF-induced osteoclastogenesis and inflam-matory bone resorption are inhibited by transcription factor RBP-J. J. Exp. Med. 209:319-334. (RTV-101)
Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY
Small GTPase Recombinant Adenoviruses
Target Name Catalog Number
Cdc42 ADV-152
Cdc42 L61 (Constitutively Active) ADV-154
Cdc42 N17 (Dominant Negative) ADV-153
Rac1 ADV-149
Rac1 L61 (Constitutively Active) ADV-151
Rac1 N17 (Dominant Negative) ADV-150
Ras N17 (Dominant Negative) ADV-145
Ras V12 (Constitutively Active) ADV-146
Ras V12C40 ADV-148
Ras V12S35 ADV-147
Rho L63 (Constitutively Active) ADV-157
Rho N19 (Dominant Negative) ADV-156
All of Cell Biolabs’ premade recombinant adenovi-ruses contain 5 x 109 viral particles per vial. They are provided as 50 µL aliquots at a concentration of 1 x 1011 viral particles/mL in TBS with 10% glycerol.
Actin Cytoskeleton Staining. Cos-7 cells were infected with purified Ad-Ras V12 (ADV-146) at 50 MOI (multiplicity of infection). Membrane ruffling was visual-ized by staining the actin cytoskeleton with Rhodamine-coupled Phalloidin.
Recent Product Citations 1. Black, S.A. et al. (2008). TGFß1 stimulates connective tissue
growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847. (ADV-145, ADV-150, ADV-153, ADV-156)
2. Mao, Y. et al. (2012). Essential diurnal Rac1 activation during retinal phagocytosis requires avß5 integrin but not tyrosine kinases focal adhesion kinase or Mer tyrosine kinase. Mol. Biol. Cell 23:1104-1114. (ADV-150)
3. Thomas, M.A. et al. (2009). E4orf1 limits the oncolytic potential of the E1B-55K-deleted adenovirus. J. Virol. 83:2406-2416. (ADV-150)
4. Rendon, B. et al. (2007). Regulation of human lung adenocarci-noma cell migration and invasion by MIF. J. Biol. Chem. 282:29910-29918. (ADV-150)
5. Yu, W.-M. et al. (2009). Laminin is required for Schwann cell morphogenesis. J. Cell Sci. 122:929-936. (ADV-150, ADV-153, ADV-154)
6. Cheng, Z.-J. et al. (2010). Co-regulation of caveolar and Cdc42-dependent fluid phase endocytosis by phosphocaveolin-1. J. Biol. Chem. 285:15119-15125. (ADV-153)
7. Perez-Moreno, M. et al. (2008). Loss of p120 catenin and links to mitotic alterations, inflammation, and skin cancer. PNAS 105:15399-15404. (ADV-156)
8. Neal, M. et al. (2013). A critical role for TLR4 induction of auto-phagy in the regulation of enterocyte migration and the patho-genesis of necrotizing enterocolitis. J. Immunol. 190:3541-3551. (ADV-156, ADV-157)
9. Vaught, D. et al. (2009). Regulation of mammary gland branch-ing morphogenesis by EphA2 receptor tyrosine kinase. Mol. Biol. Cell 20:2572-2581. (ADV-157)
10.Brantley-Sieders, D. et al. (2008). The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signal. J. Clin. Invest. 118:64-78. (ADV-157)
Target Name Vector Backbone Catalog Number
N-Ras K61 pBABEpuro RTV-222
Ras V12 pBABEpuro RTV-101
Ras V12C40 pBABEpuro RTV-104
Ras V12G37 pBABEpuro RTV-103
Ras V12S35 pBABEpuro RTV-102
RhoA L63 pBABEhygro RTV-204
Our recombinant retroviral plasmids contain a spe-cific gene cloned into a pBABE vector backbone. Each vector is supplied as 100 µL of bacterial glycerol stock.
Gene-Specific Recombinant Retroviral Vectors
Target Name Vector Backbone Catalog Number
Cdc42 L61 pBABEhygro RTV-203
myr-Rac1 pBABEpuro RTV-201
Rac1 V12 pBABEhygro RTV-202
Rac3 V12 pBABEhygro RTV-205
K-Ras pBABEpuro RTV-220
K-Ras Q61 pWZLhygro RTV-221
CELL SIGNALING & PROTEIN BIOLOGY Small GTPase/G-Protein
118 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Small GTPase Recombinant Human Proteins
Regulators of G Protein Signaling (RGS) Recombinant Human Proteins
Protein Name Tag / Location Catalog Number
Cdc42 T7 / N-term STA-700
Rab1a 6xHis / N-term STA-701
Rab1b 6xHis / N-term STA-702
Rab2a 6xHis / N-term STA-703
Rab3a 6xHis / N-term STA-704
Rab3b 6xHis / N-term STA-705
Size
50 µg
10 µg
10 µg
20 µg
20 µg
20 µg
Rab3d 6xHis / N-term 20 µg STA-706
Rab4a 6xHis / N-term 10 µg STA-707
Rab5a None 20 µg STA-708
Rab5b 6xHis / N-term 20 µg STA-709
Rab6a 6xHis / N-term 10 µg STA-710
Rab6b 6xHis / N-term 20 µg STA-711
Rab7a 6xHis / N-term 20 µg STA-712
Rab11a 6xHis / N-term 25 µg STA-713
Rab13 6xHis / N-term 10 µg STA-714
Rab14 6xHis / N-term 10 µg STA-715
Rab17 6xHis / N-term 20 µg STA-716
Rab18 T7 / N-term 10 µg STA-717
Rab21 6xHis / N-term 10 µg STA-718
Rab22 6xHis / N-term 5 µg STA-719
Rab23 6xHis / N-term 20 µg STA-720
Rab24 6xHis / N-term 20 µg STA-721
Rab27a 6xHis / N-term 25 µg STA-722
Rab27b 6xHis / N-term 20 µg STA-723
Rab32 6xHis / N-term 20 µg STA-724
Rab34 6xHis / N-term 10 µg STA-725
Protein Name Tag / Location Size Catalog Number
Rab 35 6xHis / N-term 10 µg STA-726
Rab IF 6xHis / N-term 10 µg STA-727
Rac1 6xHis / N-term 50 µg STA-728
Rac1 None 50 µg STA-729
Rac2 6xHis / N-term 20 µg STA-730
Rac3 None 20 µg STA-731
Ral A 6xHis / N-term 25 µg STA-732
Ral B 6xHis / N-term 10 µg STA-733
Ran 6xHis / N-term 10 µg STA-734
Rap1a 6xHis / N-term 10 µg STA-735
Rap1b 6xHis / N-term 10 µg STA-736
Rap2a 6xHis / N-term 10 µg STA-737
RERG 6xHis / N-term 10 µg STA-738
RHEB T7 / N-term 50 µg STA-739
RhoA 6xHis / N-term 20 µg STA-740
RhoB 6xHis / N-term 10 µg STA-741
RhoC 6xHis / N-term 10 µg STA-742
RhoD 6xHis / N-term 5 µg STA-743
RND1 6xHis / N-term 20 µg STA-744
RND3 6xHis / N-term 20 µg STA-745
SAR1A 6xHis / N-term 20 µg STA-746
H-Ras 6xHis / C-term 25 µg STA-747
K-Ras 6xHis / N-term 25 µg STA-748
N-Ras None 10 µg STA-749
R-Ras 6xHis / N-term 10 µg STA-750
R-Ras2 6xHis / N-term 50 µg STA-751
Protein Name Tag / Location Size Catalog Number
RGS2 6xHis / N-term 5 µg STA-780
RGS4 6xHis / N-term 10 µg STA-781
RGS5 6xHis / N-term 25 µg STA-782
RGS10 6xHis / N-term 25 µg STA-783
Protein Name Tag / Location Size Catalog Number
RGS16 6xHis / N-term 10 µg STA-784
RGS17 6xHis / N-term 25 µg STA-785
RGS19 6xHis / N-term 20 µg STA-786
RGS21 6xHis / N-term 20 µg STA-787
Small GTPase/G-Protein CELL SIGNALING & PROTEIN BIOLOGY
Recombinant GRP-PH Domain
Product Name Size Catalog Number
100 µg STA-200
1 mg STA-200-1MG Recombinant GRP-PH Domain
The GRP1 (general receptor for phosphoinositide) protein binds phosphatidylinositol-3,4,5-triphosphate (PIP3) through a pleckstrin homology (PH) domain and displays a region of high sequence similarity to the yeast Sec7 protein. This recombinant protein is expressed and purified from E. coli as a fusion protein, and is provided at 1.0 mg/ml in 1X PBS.
Cyclic AMP and GMP ELISA Kits
Sensitive: Detect as little as 1 pmol/mL Versatile: Suitable for use with cell and tissue lys-
ates, urine, plasma, or culture medium Convenient: Strip-well plate format with either
colorimetric or chemiluminescent detection
Cyclic AMP and cyclic GMP are important regulatory molecules in the GPCR signaling cascade. Our cAMP and cGMP ELISA Kits provide a highly sensitive method to measure low levels of cAMP or cGMP in a variety of sample types.
Product Name Detection Size Catalog Number
cAMP ELISA Kit
Colorimetric 96 Assays STA-500
5 x 96 Assays STA-500-5
Chemiluminescent 96 Assays STA-501
5 x 96 Assays STA-501-5
cGMP ELISA Kit
Colorimetric 96 Assays STA-505
5 x 96 Assays STA-505-5
96 Assays STA-506
5 x 96 Assays STA-506-5 Chemiluminescent
0
0.5
1
1.5
2
2.5
0.0 0.1 1.0 10.0 100.0 1000.0 10000.0 100000.0
cAMP (pmol/mL)
OD
45
0n
m
Standard Curve Created with the cAMP ELISA Kit, Colorimetric Format.
Recent Product Citations 1. Meyer, R. et al. (2013). GPR37 and GPR37L1 are receptors for
the neuroprotective and glioprotective factors prosaptide and prosaposin. PNAS 110:9529-9534. (STA-500)
2. Sun, Z. et al. (2011). The WD40 repeat protein WDR26 binds Gßg and promotes Gßg-dependent signal transduction and leukocyte migration. J. Biol. Chem. 286:43902-43912. (STA-500)
3. Chen, M. et al. (2010). Involvement of cAMP in nerve growth factor-triggered p35/Cdk5 activation and differentiation in PC12 cells. Am. J. Physiol. Cell Physiol. 299:C516-C527. (STA-500)
4. McCoy, K.L. et al. (2010). PAR1 and PAR2 couple to overlap-ping and distinct sets of G proteins and linked signaling path-ways to differentially regulate cell physiology. Mol. Pharmacol. 77:1005-1015. (STA-500)
5. Liu, L. et al. (2012). Radil controls neutrophil adhesion and motil-ity through ß2-integrin activation. Mol. Biol. Cell 23:4751-4765. (STA-501)
119 www.cellbiolabs.com [email protected]
120 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CELL SIGNALING & PROTEIN BIOLOGY Kinase Assays
Rho Kinase (ROCK) Activity Assays
Rho Kinase (ROCK) is a serine/threonine kinase which is a target of Rho. ROCK mediates Rho signaling and reorganizes the actin cytoskeleton via the phosphorylation of several substrates that contribute to contractility and the assembly of actin filaments.
Product Name Detection Size Catalog Number
ROCK Activity Immunoblot Kit Immunoblot 20 Assays STA-415
96-Well ROCK Activity Assay 96 Assays STA-416
5 x 96 Assays STA-416-5 Colorimetric
Results Using the ROCK Activity Immunoblot Kit. Lanes 1, 3, 5, 7: Without ROCK (negative control). Lanes 2, 4, 6, 8: With ROCK. Lanes 1 & 2: 200 ng MYPT1; Lanes 3 & 4: 100 ng; Lanes 5 & 6: 50 ng; Lanes 7 & 8: 25 ng. Phosphorylation of MYPT1 substrate was detected by anti-phospho-MYPT1 as de-scribed in the protocol.
96-Well ROCK Activity Assay Principle.
Our ROCK Activity Assays provide a non-radioactive format to measure the level of active ROCK in cell or tissue lysates. The immunoblot kit provides a conven-ient format for measuring ROCK activity in a few sam-ples, while the 96-well Activity Assay contains a strip-well plate precoated with MYPT1 for higher throughput.
Recent Product Citations 1. Georgess, D. et al. (2014). Comparative transcriptomics reveals
RhoE as a novel regulator of actin dynamics in bone-resorbing osteoclasts. Mol. Biol. 25:380-396. (STA-415)
2. Wang, J.N. et al. (2011). Response gene to complement 32 pro-motes vascular lesion formation through stimulation of smooth muscle cell proliferation and migration. Arterioscler. Thromb. Vasc. Biol. 31:e19-e26. (STA-415)
3. Xiao, L. et al. (2009). ROCK mediates phorbol ester-induced apoptosis in prostate cancer cells via p21-Cip1 upregulation and JNK. J. Biol. Chem. 284:29365-29375. (STA-415)
4. Burger, D. et al. (2011). Endothelial microparticle formation by angiotensin II is mediated via ang II receptor type I/NADPH oxi-dase/Rho kinase pathways targeted to lipid rafts. Arterioscler. Thromb. Vasc. Biol. 31:1898-1907. (STA-416)
5. Haas, B. et al. (2009). Protein kinase G controls brown fat cell differentiation and mitochondrial biogenesis. Sci. Signal. 2:ra78. (STA-416)
121 www.cellbiolabs.com [email protected]
Kinase Assays CELL SIGNALING & PROTEIN BIOLOGY
Checkpoint Kinase Activity Assays
Checkpoint kinases, including CHK1 and CHK2, can be activated in response to DNA damage prior to mi-tosis. These kinases phosphorylate Cdc25C, a pro-tein phosphatase, at Ser-216. This phosphorylation ultimately leads to cell cycle arrest, preventing mitosis and avoiding the passage of DNA damage to daugh-ter cells. Our Checkpoint Kinase Activity Assays allow you to conveniently measure the activity of CHK1 and CHK2. The assays use recombinant Cdc25C as a checkpoint kinase substrate. Phosphorylated Cdc25C (Ser216) is detected using a phospho-specific antibody. Checkpoint Kinase Activitiy As-says are available in two formats: a Western blot assay and a 96-well plate-based activity assay.
Product Name Detection Size Catalog Number
Checkpoint Kinase Activity Immunoblot Kit Immunoblot 20 Assays STA-413
96-Well Checkpoint Kinase Activity Assay Kit 96 Assays STA-414
5 x 96 Assays STA-414-5 Colorimetric
96-Well Checkpoint Kinase Activity Assay Principle.
CHK1 Activity Using the Checkpoint Kinase Activity Immunoblot Kit. Lane 1: Negative Control; Lane 2: 10 ng of active CHK1.
122
GFP Quantitation Kit
Product Name Detection Size Catalog Number
GFP Quantitation Kit Fluorometric 100 Assays AKR-120
Most imaging studies of rGFP are qualitative, and quantitation by FACS is time consuming and expen-sive. Our GFP ELISA kit uses a standard microplate reader to quantify GFP levels with extremely high sensitivity. This kit will detect GFP form Aequorea victoria as well as its variants.
Standard Curve Generated with the GFP ELISA Kit.
Sensitive: Detection limit of 30 pg/mL Versatile: Quantify GFP and its variants: BFP,
CFP or YFP Easy Quantitation: Measure GFP levels in a
standard microplate reader
Recent Product Citations 1. Mango, R. et al. (2014). C-C chemokine receptor 5 on pulmo-
nary mesenchymal cells promotes experimental metastasis via the induction of erythroid differentiation regulator 1. Mol. Cancer Res. 12:274-282.
2. Mitchell, A. et al. (2014). Promyelocytic leukemia protein is a cell-intrinsic factor inhibiting parvovirus DNA replication. J. Virol. 88:925-936.
3. Coghill, J.M. et al. (2013). CC chemokine receptor 8 potentiates donor Treg survival and is critical for the prevention of murine graft-versus-host disease. Blood 122:825-836.
4. Pedersen, J. et al. (2012). Glucose metabolism is altered after loss of L cells and -cells but not influenced by loss of K cells. Am. J. Physiol. Endocrinol. Metab. 304:E60-E73.
5. Zhou, W. et al. (2012). Inducible podocyte injury and proteinuria in transgenic Zebrafish. J. Am. Soc. Nephrol. 23:1039-1047.
6. Fonseca, J.P. et al. (2012). In vivo polycomb kinetics and mitotic chromatin binding distinguish stem cells from differentiated cells. Genes and Dev. 26: 857-871.
7. Coghill, J.M. et al. (2010). Separation of graft-versus-host dis-ease from graft-versus-leukemia responses by targeting CC-chemokine receptor 7 on donor T cells. Blood 115:4914-4922.
8. Rajan, S. et al. (2010). In vitro processing and secretion of mu-tant insulin proteins that cause permanent neonatal diabetes.
Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules
We offer a variety of tools for various reporter molecules:
Reporter Assays Reporter Stable Cell Lines Recombinant Fluorescent Proteins
Antibodies to Reporter Molecules Reporter Viral Vectors
Reporter Assays, Cell Lines and Reagents
GFP ELISA Kit
Recent Product Citations 1. Pfeiffer, B. et al. (2012). Using translational enhancers to in-
crease transgene expression in Drosophila. PNAS 109:6626-6631.
2. Wamboldt, Y. et al. (2009). Participation of leaky ribosome scan-ning in protein dual targeting by alternative translation initiation in higher plants. Plant Cell 21:157-167.
Product Name Detection Size Catalog Number
GFP ELISA Kit Colorimetric 96 Assays AKR-121
When direct quantitation of GFP fluorescence levels is desired, our GFP Quantitation Kit provides a su-perior method over time-consuming flow cytometry and semi-quantitative imaging techniques. This kit measures fluorescence levels directly in a plate-based fluorometer.
123
ß-Galactosidase Staining Kit
LacZ is a commonly used reporter gene in transfec-tion experiments because its gene product, ß-galactosidase, is extremely stable and resistant to proteolytic degradation, making it easy to assay. Our ß-Galactosidase Staining Kit provides an effi-cient, easy-to-use method to determine the transfec-tion efficiency of the LacZ gene.
Product Name Size Catalog Number
ß-Galactosidase Staining Kit 75 Assays AKR-100
Cell Line Catalog Number
293/CFP AKR-270
293/GFP AKR-200
293/Luc AKR-230
Reporter Stable Cell Lines
Recent Product Citation Black, S.A. et al. (2008). TGFß1 stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibro-blasts through a RhoA-independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFß1-induced CCN2/CTGF expression. J. Biol. Chem. 283:10835-10847.
www.cellbiolabs.com [email protected]
Reporter Molecules CELL SIGNALING & PROTEIN BIOLOGY
Cell Line Catalog Number
A549/GFP AKR-209
BT-549/GFP AKR-255
ES-2/GFP AKR-206
HeLa/GFP AKR-213
MCF-7/GFP AKR-211
MCF-7/Luc AKR-234
MDA-MB-231/GFP AKR-201
MDA-MB-231/GFP-RFP AKR-221
MDA-MB-231/Luc AKR-231
293/YFP AKR-280
293T/GFP-Puro AKR-202
Cell Line Catalog Number
MDA-MB-468/GFP AKR-204
NIH3T3/GFP AKR-214
OVCA429/GFP AKR-212
OVCAR-5/RFP AKR-254
SKOV-3/GFP-Luc AKR-225
SKOV-3/Luc AKR-232
SKOV-3/RFP AKR-253
T47D/GFP AKR-208
MDA-MB-436/GFP AKR-203
MDA-MB-436/RFP AKR-252
MDA-MB-231/RFP AKR-251
RFP ELISA Kit
Recent Product Citations 1. Zhang, K. et al. (2014). Block-cell-
printing for live single-cell printing. PNAS 111:2948-2953. (AKR-201, AKR-211, AKR-252)
2. Zhang, W. et al. (2012). Microfluidics separation reveals the stem-cell-like deformability of tumor-initiating cells. PNAS 109:18707-18712. (AKR-211, AKR-252)
3. Weerakkody, D. et al. (2013). Family of pH (low) insertion peptides for tumor targeting. PNAS 110:5834-5839. (AKR-213)
Each cell line expresses one or more reporter molecules.
Product Name Detection Size Catalog Number
RFP ELISA Kit Colorimetric 96 Assays AKR-122
Sensitive: Detection limit of 150 pg/mL
Easy Quantification: Measure RFP levels in a standard 96-well microplate reader
Our RFP ELISA Kit provides a convenient, sensitive alternative to imaging systems and time-consuming FACS quantitation. The assay quantifies a wide vari-ety of red fluorescent protein variants including DsRed, TagRFP, TurboRFP, tdTomato, mCherry, mKate, mRuby, mBanana, mOrange, mPlum, and mStrawberry.
X-Gal Staining of Infected HUVEC Cells and Chick CAM Tissue. Left: HUVEC cells were infected with purified Ad-ß-Gal at 50 MOI (multiplicity of infection). X-gal staining was performed after 48 hour infection period. Right: Purified Ad-ß-Gal was injected intravenously into a 10-day old chick embryo. After three days, X-gal staining was performed on the chick chorioallanoic membrane tissue.
CELL SIGNALING & PROTEIN BIOLOGY Reporter Molecules
124 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Monoclonal Antibodies to Reporter Molecules
Antibodies are provided at a concentration of 1 mg/mL. GFP antibody also recognizes EGFP, YFP, EYFP and CFP. RFP antibody recognizes Tag-RFP, Turbo-RFP, DeRed, mCherry and mOrange. Antibodies are suitable for Western blot, Immunostaining, ELISA and Dot blot.
Size Catalog Number
Mouse Anti-GFP Monoclonal Antibody (clone GF28R) 100 µg AKR-020
Mouse Anti-RFP Monoclonal Antibody (clone RF5R) 100 µg AKR-021
Product Name
Reporter Viral Vectors
Premade viruses and viral constructs with reporter molecules make ideal controls for viral gene delivery experi-ments.
Recombinant Fluorescent Proteins
Product Name Size Catalog Number
Recombinant EGFP 100 µg STA-201
5 x 100 µg STA-201-5
100 µg STA-202
5 x 100 µg STA-202-5 Recombinant RFP
Recombinant EGFP and RFP are provided at a con-centration of 1 mg/mL and includes a 6xHis-tag at the C-terminus.
Recent Product Citation Sokolova, E. et al. (2013). Enhanced transcription rates in mem-brane-free protocells formed by coacervation of cell lysate. PNAS 110:11692-11697. (STA-201)
Recent Product Citation Maamary, J. et al. (2012). Attenuated influenza virus constructs with enhanced hemagglutinin protein expression. J. Virol. 86:5782-5790. (AKR-021)
Reporter AAV Reporter Adenoviruses
Reporter Lentiviruses
Reporter Retroviral Plasmids
Product Name Catalog Number
AAV1-GFP Control Virus AAV-301
AAV2-Cre Control Virus AAV-310
AAV2-Luc Control Virus AAV-320
AAV3-GFP Control Virus AAV-303
AAV5-GFP Control Virus AAV-305
AAV6-GFP Control Virus AAV-306
AAV2-GFP Control Virus AAV-302
Product Name Catalog Number
GFP Lentivirus Control LTV-300
RFP Lentivirus Control LTV-301
Product Name Catalog Number
-Galactosidase ADV-002
Firefly Luciferase ADV-008
GFP ADV-004
Target Name Catalog Number
GFP RTV-002
GFP RTV-051
Vector Backbone
pBABE
pMCs
GFP pMX RTV-050
GFP pMYs RTV-052
GFP-Puro pMX RTV-053
Epitope Tags/Antibodies CELL SIGNALING & PROTEIN BIOLOGY
125 www.cellbiolabs.com [email protected]
Size Catalog Number
Mouse Anti-FLAG Tag Monoclonal Antibody (clone FG4R) 100 µg AKR-004
Mouse Anti-GST Tag Monoclonal Antibody (clone GST.B6) 100 µg AKR-005
Product Name
Mouse Anti-HA Tag Monoclonal Antibody (clone HA.C5) 100 µg AKR-006
Mouse Anti-His Tag Monoclonal Antibody (clone HIS.H8) 100 µg AKR-003
Mouse Anti-Myc Tag Monoclonal Antibody (clone Myc.A7) 100 µg AKR-007
Mouse Anti-V5 Tag Monoclonal Antibody (clone E10) 100 µg AKR-008
Mouse Anti-GAPDH Monoclonal Antibody 100 µg AKR-001
Mouse Anti-ß-Actin Monoclonal Antibody 100 µg AKR-002
Mouse Anti-ß-Tubulln Monoclonal Antibody 100 µg AKR-009
Monoclonal Antibodies to Epitope Tags
Antibodies are provided at a concentration of 1 mg/mL. GAPDH, ß-Actin and ß-Tubulin are also available as loading controls. All are suitable for Western blot, Immunostaining, ELISA, Immunoprecipitation, and Dot blot.
His-Tag Protein ELISA Kit
Our His-Tag Protein ELISA Kit allows you to detect and quantify His-tagged protein samples simply and reliably by comparing unknown samples with a recombinant standard. The kit is suitable for use with cell lysates and tissue homogenates.
Sensitive: Detect as little as 1 ng/mL protein or 50 pM of 6xHis-tag residues
Versatile: Use with proteins containing His-tag at either N- or C-terminus
Rapid Antibody Purification Kit
Product Name Size Catalog Number
Rapid Antibody Purification Kit 10 Preps AKR-160
Species mg of IgG per Prep
Bovine 15-20
Goat 6-12
Human 20-30
Mouse 6-12
Rabbit 15-20
Our Rapid Antibody Purification Kit is designed for fast, single-step purification of high-quality IgG from ascites, serum, tissue culture media or hybridoma supernatants. IgG-containing samples are incubated with immobilized Protein A in the presence of a bind-ing buffer. Non-IgG components are washed and IgG is subsequently eluted. The supplied Protein A col-umn is suitable for 10 purification preps. The capac-ity per prep depends on the species of IgG; exam-ples are listed in the column at right. Capacity per Prep for the Rapid Antibody Purification Kit.
Recent Product Citation Dong, Y. et al. (2013). HMGB1 protein does not mediate the in-flammatory response in spontaneous spinal cord regeneration: A hint for CNS regeneration. J. Biol. Chem. 288:18204-18218.
Product Name Detection Size Catalog Number
His-Tag Protein ELISA Kit Colorimetric 96 Assays AKR-130
PhosphoBLOCKER™ Reagent
Dry Milk
PhosphoBLOCKER™ Western Blot Blocking Reagent
Superior Blocking with PhosphoBLOCKER™ Reagent. A549 cell lysate was blocked with dry milk or PhosphoBLOCKER before detection with anti-Phospho-p38 antibody.
Product Name Size Catalog Number
1 L AKR-103
4 L AKR-104 PhosphoBLOCKER™ Western Blot Blocking Reagent
Product Name Size Catalog Number
Phospho Antibody Stripping Solution 10 mL AKR-102
Phospho Antibody Stripping Solution
Western blot blockers such as dry milk or serum are sufficient to block unreactive sites on the membrane. However, they are not designed to preserve phos-phoprotein antigens during blotting.
High Sensitivity: Enhances low level phospho-protein signal without increasing background
Easy-to-use: Premixed dry blend
This solution removes anti-phosphoantibodies selec-tively from blots without significantly affecting the immobilized proteins, allowing re-probing of the blot with the same or a different antibody. Stripping of antibodies is done at room temperature, so no heat-ing of blots is required.
Multiple Blotting and Stripping of 4G10 Phosphotyrosine Antibody.
Recent Product Citations 1. Marques, J. et al. (2013). CRMP2 tethers kainate receptor activ-
ity to cytoskeleton dynamics during neuronal maturation. J. Neu-rosci. 33:18298-18310.
2. Ferreira, E. et al. (2013). Inflammatory cytokines induce a unique mineralizing phenotype in mesenchymal stem cells de-rived from human bone marrow. J. Biol. Chem. 288:29550-29561.
3. Rosich, L. et al. (2012). Counteracting autophagy overcomes resistance to everolimus in mantle cell lymphoma. Clin. Cancer Res. 18:5278-5289.
4. Martinez, P. et al. (2012). 53BP1 deficiency combined with te-lomere dysfunction activates ATR-dependent DNA damage response. J. Cell Biol. 197:283-300.
CELL SIGNALING & PROTEIN BIOLOGY Protein Phosphorylation
126 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Product Name Size Catalog Number
2 preps AKR-105
5 preps AKR-106 Phosphoprotein Purification Kit
Phosphoprotein Purification Kit
Enrichment of p-ERK. HeLa cell lysate was incubated with the Phos-phoprotein Enrichment Matrix from the Phos-phoprotein Purification Kit.
Our Phosphoprotein Purification Kit allows you to enrich your phosphoprotein samples quickly and easily. Phosphorylated proteins are affinity purified from lysates with a single-step purification / enrich-ment matrix. The entire procedure takes about 4 hours. Each prep can process 2.5 mg of total lysate protein, or approximately one confluent 10 cm dish.
Our Bacterial Protein Extraction Reagents contain a gentle, non-ionic detergent formulation that quickly extracts functional recombinant proteins from E. coli without mechanical disruption. The reagents are compatible with 6xHis and GST affinity purification systems and are suitable for small-scale or large-scale protein extraction.
127 www.cellbiolabs.com [email protected]
Protein Isolation CELL SIGNALING & PROTEIN BIOLOGY
Rapid GST Inclusion Body Solubilization and Renaturation Kit
Product Name Size Catalog Number
Rapid GST Inclusion Body Solubilization and Renaturation Kit 1 Kit AKR-110
The Rapid GST Inclusion Body Solubilization and Re-naturation Kit is designed to retrieve expressed GST fusion proteins in soluble form after lysis and extrac-tion. Each kit contains sufficient reagents to solubilize and renature up to 5-10 liters of bacterial culture.
Fast Results: No lengthy dialysis or dilution steps Easy-to-Use: No pH variation or redox pair Optimized: Designed specifically to solubilize and
renature GST inclusion bodies
Solubilization and Renaturation of GST-RTK Fusion Protein. Lane 1: MW STD; Lane 2: Whole E.Coli lysate; Lane 3, 7, 11: No detergent; Lane 4, 8, 12: 32-fold dilution; Lane 5, 9, 13: 8-fold dilution; Lane 6, 10, 14: 2-fold dilution.
1 2 3 4 5 6 7 8 9 10 11 12 13 14
Pre Beads Post Beads GS Beads
Recent Product Citations 1. Keller, D. et al. (2014). Mechanisms of HsSAS-6 assembly pro-
moting centriole formation in human cells. J. Cell Biol. 204:697-712.
2. Lalani, S. et al. (2013). MCTP2 is a dosage-sensitive gene re-quired for cardiac outflow tract development. Hum. Mol. Genet. 10.1093/hmg/ddt283.
Nuclear/Cytosolic Fractionation Kit
Product Name Size Catalog Number
20 preps AKR-171
100 preps AKR-172 Nuclear/Cytosolic Fractionation Kit
The Nuclear/Cytosolic Fractionation Kit provides a simple and fast tool to isolate nuclear extract from the cytoplasmic fraction of mammalian cells. The opti-mized protocol provides high protein recovery and low cross-contamination in less than 2 hours. Ex-tracted fractions are functional and suitable for down-stream assays such as DNA footprinting, pre-mRNA splicing, gel shift assays, reporter assays, enzyme activity assays, and Western blot.
Size Catalog Number
5X Bacterial Protein Extraction Reagent (Phosphate) 50 mL AKR-181
5X Bacterial Protein Extraction Reagent (Tris) 50 mL AKR-180
Product Name
Bacterial Protein Extraction Reagents
HEK293 Cell Fractionation. Whole cell (W), cytosolic (C), and nuclear (N) frac-tions were im-munoblotted with Anti-Tubulin (cytosol specific) or Anti-Lamin A/C (nuclear specific).
CELL SIGNALING & PROTEIN BIOLOGY Protein Quantitation
128 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
High Sensitivity Protein Quantitation Assay Kit
Our High Sensitivity Protein Quantitation Assay Kit provides a robust method to quantify protein concen-trations in the µg/mL range. The fluorometric format creates a significant sensitivity advantage over stan-dard colorimetric methods such as Bradford or BCA assays.
0
50
100
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250
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BSA Standard (ug/mL)
RF
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0
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BSA Standard (ug/mL)
RF
U
Highly Sensitive: Quantify protein concentrations as low as 5 µg/mL
Fast: Obtain results in 5 to 15 minutes Easy-to-use: Read on a 96-well fluorescence
plate reader
Standard Curve Generated with the High Sensitivity Protein Quantitation Assay Kit.
Product Name Size Catalog Number
High Sensitivity Protein Quantitation Assay Kit 100 Assays AKR-185
RIPA Buffer
RIPA buffer is a popular reagent for lysis of both adherent and suspension cells in culture, as well as making tissue homogenates. RIPA buffer extracts cytoplasmic, membrane, and nuclear proteins for a variety of down-stream protein assays and immunoassays. Our RIPA Buffer is provided as a 5X concentrate and is available with or without a Protease Inhibitor Cocktail.
Product Name Size Catalog Number
5X RIPA Buffer 20 mL AKR-191
5X RIPA Buffer, with Protease Inhibitor Cocktail 20 mL AKR-190
METABOLISM RESEARCH Lipoprotein Metabolism
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Lipoproteins are important assemblies of proteins and lipids that have implications in car-diovascular and related diseases. Our kits and reagents can help streamline the study of various members of lipoprotein metabolism pathways:
Lipoprotein Metabolism Research
Cholesterol / Lipoproteins Apolipoproteins Oxidized LDL Lipoprotein Receptors Cholesteryl Ester Pathway
Lipoprotein Lipase Triglycerides Free Fatty Acids Phospholipids Serum Proteins
Total Cholesterol Assay Kits
Product Name Detection Size Catalog Number
Colorimetric 192 Assays STA-384
Fluorometric 192 Assays STA-390 Total Cholesterol Assay Kit
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Cholesterol Values from Normal Human Serum and Plasma Samples. Relative Fluorescence Unit (RFU) values are then com-pared against a Cholesterol Standard Curve (not shown).
Sensitive: Detect as little as 100 nM Fast: Simple 30 minute protocol Easy-to-use: Detect in a 96-well plate reader;
colorimetric and fluorometric formats available
Assay Principle for the Total Cholesterol Assay Kit (Fluorometric).
Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester, which is the predominant form of cholesterol transport and stor-age. Our Total Cholesterol Assay Kits provide a simple plate-based format that measures the amount of cho-lesterol in serum, plasma, cell lysates or tissue ho-mogenates. In the presence of cholesterol esterase, the assay will measure total cholesterol in both forms. In the absence of the esterase, the assay will meas-ure only free cholesterol. Quantitation is performed in a 96-well plate reader with your choice of colorimetric or fluorescence-based detection.
Recent Product Citations 1. Marino, A. et al. (2014). ITCH deficiency protects from diet-
induced obesity. Diabetes 63:550-561. (STA-384) 2. Mathews, E. et al. (2014). Mutation of 3-hydroxy-3-
methylglutaryl CoA synthase I reveals requirements for isopre-noid and cholesterol synthesis in oligodendrocyte migration arrest, axon wrapping, and myelin gene expression. J. Neurosci. 34:3402-3412. (STA-384)
Lipoprotein Metabolism METABOLISM RESEARCH
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HDL and LDL/VLDL Cholesterol Assay Kit
Product Name Detection Size Catalog Number
HDL and LDL/VLDL Cholesterol Assay Kit Fluorometric 192 Assays STA-391
Our HDL and LDL/VLDL Cholesterol Assay Kit is similar in principle to our Total Cholesterol Assay Kit, but allows you to quantify HDL and LDL/VLDL sepa-rately in serum samples. After separating samples into HDL and LDL/VLDL fractions, the fluorometric assay is run according to the Assay Principle for the Total Cholesterol Assay Kit (previous page). In the presence of cholesterol esterase, the assay will measure total cholesterol in both free cholesterol and cholesteryl ester forms. In the absence of the es-terase, the assay will measure only free cholesterol. Quantitation of cholesteryl ester alone may be calcu-lated by subtracting free cholesterol from total choles-terol levels. Quantitation of Total Cholesterol, LDL/VLDL, and HDL.
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HDL-Cholesterol Assay Kit
Human Lipoproteins
Product Name Size Catalog Number
Human High Density Lipoprotein (HDL) 100 µg STA-243
Human High Density Lipoprotein-2 (HDL-2) 100 µg STA-244
Human High Density Lipoprotein-3 (HDL-3) 100 µg STA-245
Human Low Density Lipoprotein (LDL) 100 µg STA-241
Human Low Density Lipoprotein (LDL), Copper (Cu++) Oxidized 100 µg STA-214
Human Low Density Lipoprotein (LDL), Malondialdehyde Modified 100 µg STA-212
Human Low Density Lipoprotein (LDL), Nitrated 100 µg STA-213
Human Very Low Density Lipoprotein (VLDL) 100 µg STA-242
Product Name Detection Size Catalog Number
HDL-Cholesterol Assay Kit Fluorometric 96 Assays STA-394
Sensitive: Detect as little as 1 µM Fast: Simple 45 minute assay protocol Easy-to-use: Detect in a 96-well plate fluores-
cence-based plate reader
Our HDL-Cholesterol Assay Kit provides a simple plate-based format similar to our Total Cholesterol Assays (previous page). This kit measures the amount of cholesterol in the HDL fraction isolated from serum, plasma, cell lysates or tissue homoge-nates.
Our human lipoproteins are isolated from human plasma following ultracentrifugation.
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Product Name Detection Size Catalog Number
Human ApoAI ELISA Kit Colorimetric 96 Assays STA-362
Human ApoAII ELISA Kit Colorimetric 96 Assays STA-363
Human ApoCI ELISA Kit Colorimetric 96 Assays STA-364
Human ApoCII ELISA Kit Colorimetric 96 Assays STA-365
Human ApoCIII ELISA Kit Colorimetric 96 Assays STA-366
Human ApoE ELISA Kit Colorimetric 96 Assays STA-367
Human ApoB ELISA Kit Colorimetric 96 Assays STA-368
Human Apo(a) ELISA Kit Colorimetric 96 Assays STA-359
Human Apolipoprotein ELISA Kits
Apolipoproteins comprise the protein component of lipoprotein assemblies. Apolipoproteins fall into two classes: Non-exchangeable Apo B associates with LDL Exchangeable Apo A, C and E associate with
HDL Our Human Apo ELISA Kits provide a convenient and sensitive method for quantifying specific apolipopro-teins in serum, plasma, or other biological fluids.
Human ApoB ELISA Standard Curve. Detection Limits of Cell Biolabs’ Human Apo ELISA Kits.
Apo AI
Apo AII
Apo B
Apo CI
Apo CII
Apo CIII
Apo E
50 pg/mL
0.3 ng/mL
1 ng/mL
200 pg/mL
1 ng/mL
50 pg/mL
200 pg/mL
Apo (a)
1 ng/mL
Human Apolipoproteins
Product Name Size Catalog Number
Human Albumin, Malondialdehyde Modified 100 µg STA-210
Human Apolipoprotein AI 100 µg STA-232
Human Apolipoprotein AII 100 µg STA-233
Human Apolipoprotein B-100 100 µg STA-234
Human Apolipoprotein B-100, Malondialdehyde Modified 100 µg STA-211
Human Apolipoprotein CI 100 µg STA-235
Human Apolipoprotein CIII 100 µg STA-237
Following ultracentrifugation, lipoproteins are isolated from human plasma. Water-soluble apolipoproteins are then purified from delipidated lipoprotein.
Lipoprotein Metabolism METABOLISM RESEARCH
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Human ApoAI and ApoB Duplex ELISA Kit
Product Name Detection Size Catalog Number
Human ApoAI and ApoB Duplex ELISA Kit Colorimetric 96 Assays STA-361
Efficient: Quantify two apolipoproteins from the same sample in just a few hours
Sensitive: Detect as little as 0.1 ng/mL of ApoAI and 1 ng/mL of ApoB from serum or plasma
Quantitative: Compare results to known ApoAI and ApoB standards
Human ApoAI and ApoB Duplex ELISA Kit Assay Principle.
As the primary protein components of HDL and LDL respectively, ApoAI and ApoB are arguably the most significant apolipoproteins in lipid metabolism re-search. Our Human ApoAI and ApoB Duplex ELISA Kit pro-vides a convenient tool to quantify both proteins in a single serum or plasma sample. Unlike other multi-plex assays, our ApoAI and ApoB Duplex ELISA does not require a luminometer for detection. Simply quantify both proteins using a standard colorimetric ELISA plate reader. The ELISA plate is coated with Anti-ApoAI and Anti-ApoB antibodies, which respectively capture HDL and LDL from the sample. Biotinylated Anti-ApoAI and Anti-ApoB detection antibodies are added, followed by enzyme conjugates. Alkaline phosphatase sub-strate is added allowing quantitation of ApoAI. After washing, HRP is added to allow quantiation of ApoB.
Product Name Detection Size Catalog Number
Sheep Anti-Human Apolipoprotein (a) Polyclonal Antibody Immunoblot/ELISA 100 µg STA-131
Goat Anti-Human Apolipoprotein AI Polyclonal Antibody Immunoblot/ELISA 100 µg STA-132
Rabbit Anti-Human Apolipoprotein AII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-133
Goat Anti-Human Apolipoprotein B-100/48 Polyclonal Antibody Immunoblot/ELISA 100 µg STA-134
Rabbit Anti-Human Apolipoprotein CI Polyclonal Antibody Immunoblot/ELISA 100 µg STA-135
Rabbit Anti-Human Apolipoprotein CII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-136
Rabbit Anti-Human Apolipoprotein CIII Polyclonal Antibody Immunoblot/ELISA 100 µg STA-137
Goat Anti-Human Apolipoprotein E Polyclonal Antibody Immunoblot/ELISA 100 µg STA-138
Antibodies to Apolipoproteins
Antibodies are affinity purified.
METABOLISM RESEARCH Lipoprotein Metabolism
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OxiSelect™ Human Oxidized LDL ELISA Kits
LDL contains a hydrophobic core of various lipids surrounded by one molecule of Apolipoprotein B-100 (ApoB-100), which promotes solubility of the LDL in blood. LDL, often described as “bad” cholesterol, is even more dangerous when it becomes oxidized. Oxi-dized LDL (OxLDL) is more reactive with surrounding tissues and can collect within the inner lining of arter-ies. Our OxiSelect™ Human Oxidized LDL ELISA Kits are designed for the detection and quantitation of modi-fied LDL in human plasma or serum. Kits are avail-able to detect MDA-LDL, CML-LDL, or HNE-LDL in either the protein or lipid component of LDL. Our OxPL-LDL kit specifically detects oxidation in the phospholipid component of LDL.
Product Name Detection Size Catalog Number
OxiSelect™ Human Oxidized LDL ELISA Kit (CML-LDL Quantitation) Colorimetric 96 Assays STA-388
OxiSelect™ Human Oxidized LDL ELISA Kit (HNE-LDL Quantitation) Colorimetric 96 Assays STA-389
OxiSelect™ Human Oxidized LDL ELISA Kit (MDA-LDL Quantitation) Colorimetric 96 Assays STA-369
OxiSelect™ Human Oxidized LDL ELISA Kit (OxPL-LDL Quantitation) Colorimetric 96 Assays STA-358
Sensitive: Detect as little as 50 ng/mL of MDA-LDL, 150 ng/mL of CML-LDL, 150 ng/mL of HNE-LDL, or 100 ng/mL of OxPL-LDL
Quantitative: Compare unknown samples with provided copper oxidized LDL standard
Quantitation of MDA-LDL in Serum and Plasma Samples. Se-rum and plasma samples were treated with LDL Precipitation Solu-tion. Precipitated LDL pellets were resuspended in 1.6 mL of PBS before further dilution 1:160 in Assay Diluent according to the As-say Protocol. OxiSelect™ Human Oxidized LDL ELISA Assay Principle.
MDA is the most commonly found damage marker in oxidized LDL, but it can degrade in frozen samples after 1-2 months. CML and HNE, while less commonly found in OxLDL, may be more reliably detectable in samples that have been frozen for several months.
Lipoprotein Metabolism METABOLISM RESEARCH
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Human LDL Receptor ELISA Kit
Human LOX-1 ELISA Kit
Product Name Detection Size Catalog Number
Human LDLR ELISA Kit Colorimetric 96 Assays STA-386
Product Name Detection Size Catalog Number
Human LOX-1 ELISA Kit Colorimetric 96 Assays STA-387
Recent Product Citation Wu, J. et al. (2013). Clinical nephrology—IgA nephropathy, lupus nephritis, vasculitis. Nephrol. Dial. Transplant. 28:i175-i184.
Cholesterol can be toxic when accumulated in excess in cell membranes. The low-density lipoprotein recep-tor (LDLR) is the primary means of removing choles-terol from the circulation. LDLR is a transmembrane protein that transports cholesterol-carrying lipoprotein particles (primarily LDL) into cells. Receptor-ligand complexes enter the cell by endocytosis; bound lipo-proteins are subsequently released in the low-pH set-ting of the endosome, while the receptors return to the cell surface. Our Human LDLR ELISA Kit provides a simple, con-venient method for the detection and quantitation of LDL receptor in a variety of human sample types.
Sensitive: Detect as little as 50 pg/mL of human LDLR
Versatile: Assay is compatible with plasma, serum, cell lysates and tissue homogenates
Quantitative: Compare results to a known human LDLR standard
Standard Curve Generated with the Human LDLR ELISA Kit.
Standard Curve Generated with the Human LOX-1 ELISA Kit.
Monocytes and macrophages can form atheroscle-rotic lesions when they take in oxidized LDL (OxLDL). Uptake is done via the lectin-like oxidized LDL recep-tor-1 (LOX-1), which is expressed in vascular endo-thelium as well as in vascular smooth muscle cells, differentiated macrophages and platelets. LOX-1 can be cleaved and released as a soluble form (sLOX-1), which can serve as a prognostic biomarker in serum for early acute coronary syndromes, stroke and coro-nary heart disease. Our Human LOX-1 ELISA Kit provides a simple, con-venient method for the detection and quantitation of LOX-1 receptor in a variety of sample types.
Sensitive: Detect as little as 40 pg/mL of human LOX-1
Versatile: Assay is compatible with plasma, serum, cell lysates, tissue homogenates, or cell culture supernatants
Quantitative: Compare results to a known human LOX-1 standard
CETP Promotes Bidirectional Transfer of Cholesteryl Esters (CE) and Triglycerides (TG) Between Lipoproteins.
METABOLISM RESEARCH Lipoprotein Metabolism
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Human PCSK9 ELISA Kit
Product Name Detection Size Catalog Number
Human PCSK9 ELISA Kit Colorimetric 96 Assays STA-385
Standard Curve Generated with the Human PCSK9 ELISA Kit.
Proprotein convertase subtilisin kexin 9 (PCSK9) is a member of the proteinase K subfamiliy of subtilisin-related serine endoproteases. PCKS9 mediates LDL receptor (LDLR) degradation by binding to the EGF domain of the LDLR. This binding prevents LDLR from being sorted to the endosomes for recycling back to the cell surface. Instead, the PCSK9/LDLR complex is distributed to the lysosomes for degrada-tion. Our Human PCKS9 ELISA Kit provides a simple, convenient method for the detection and quantitation of PCSK9 in a variety of human sample types.
Sensitive: Detect as little as 150 pg/mL of human PCSK9
Versatile: Assay is compatible with plasma, serum, and cell and tissue lysates
Quantitative: Compare results to a known human PCSK9 standard
Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit
Product Name Detection Size Catalog Number
Human Cholesteryl Ester Transfer Protein (CETP) ELISA Kit Colorimetric 96 Assays STA-614
Cholesterol exists within lipoproteins in two forms: a free alcohol and a fatty cholesteryl ester. The choles-teryl ester is the predominant form of cholesterol transport and storage. Cholesteryl ester transfer protein (CETP) promotes the transfer of both cholesteryl esters and triglyc-erides between various types of lipoprotein particles: HDL, LDL, VLDL, and chylomicrons. Our Human CETP ELISA Kit provides a simple, con-venient method for the detection and quantitation in a variety of human sample types.
Sensitive: Detect as little as 60 ng/mL of human CETP
Versatile: Assay is compatible with plasma, serum, and other biological fluids
Quantitative: Compare results to a known human CETP standard
Lipoprotein Metabolism METABOLISM RESEARCH
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Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit
Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit
Product Name Detection Size Catalog Number
Lecithin Cholesterol Acyltransferase (LCAT) Activity Assay Kit Fluorometric 100 Assays STA-615
Product Name Detection Size Catalog Number
Lecithin Cholesterol Acyltransferase (LCAT) ELISA Kit Colorimetric 96 Assays STA-616
Lecithin cholesterol acyltransferase (LCAT) is an en-zyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cell-associated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipopro-teins in the blood, including HDL and LDL. Our LCAT Activity Assay Kit provides a simple, con-venient method for measuring the phospholipase ac-tivity of LCAT in a variety of sample types including plasma, serum, cell lysates and tissue homogenates. Quantitation of LCAT activity is performed in a 96-well fluorescence-based plate reader. This assay may also be used to quantify other cal-cium independent phospholipase activities such as lipoprotein phospholipase A2 (LP-PLA2).
Assay Principle for the LCAT Activity Assay Kit. The close proximity of fluorescence labels on a dual-labeled fluorogenic probe keeps the fluorescence quenched. Upon cleavage of the probe by LCAT, fluorescence of the monomers can be measured at an excitation of 342 nm and emission of 400 nm.
Standard Curve Generated with the Lecithin Cholesterol Acyl-transferase (LCAT) ELISA Kit.
Lecithin cholesterol acyltransferase (LCAT) is an en-zyme that is associated with lipoproteins and plays a key role in promoting the transfer of excess cell-associated cholesterol from peripheral tissues to the liver to be excreted. LCAT catalyzes the transfer of an sn-2 acyl group from phosphatidylcholine to cholesterol, forming a cholesteryl ester. LCAT is bound to various lipopro-teins in the blood, including HDL and LDL. Our LCAT ELISA Assay Kit provides a simple, con-venient method for quantifying LCAT levels in a vari-ety of sample types including plasma, serum, and cell and tissue lysates. Quantitation of LCAT activity is performed in a standard 96-well plate reader.
Sensitive: Detect as little as 30 ng/mL of LCAT Versatile: Suitable for human, mouse, rat or rabbit
samples
METABOLISM RESEARCH Lipoprotein Metabolism
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Lipoprotein Lipase (LPL) Activity Assay Kit
Our Lipoprotein Lipase (LPL) Activity Assay Kit pro-vides a simple, convenient method to measure LPL activity in a variety of sample types. This kit uses a fluorogenic triglyceride analog as a lipase substrate. The quenched substrate is cleaved at the sn-1 posi-tion by LPL producing a fluorescent product that can be detected in a 96-well fluorescence plate reader. This assay will also measure the activity of endothe-lial and hepatic lipases. It cannot distinguish between these lipases and LPL.
Lipoprotein Lipase (LPL) ELISA Kit
Recent Product Citations 1. Dib, L. et al. (2014). LXRalpha fuels fatty acid-stimulated oxygen
consumption in white adipocytes. J. Lipid Res. 55:247-257. 2. Navab, M. et al. (2013). Transgenic 6F tomatoes act on the
small intestine to prevent systemic inflammation and dyslipide-mia caused by Western diet and intestinally derived lysophos-phatidic acid. J. Lipid Res. 54:3403-3418.
Product Name Detection Size Catalog Number
Lipoprotein Lipase (LPL) Activity Assay Kit Fluorometric 100 Assays STA-610
Product Name Detection Size Catalog Number
Lipoprotein Lipase (LPL) ELISA Kit Colorimetric 96 Assays STA-611
Assay Principle for the LPL Activity Assay Kit. The fluorogenic substrate is initially quenched and non-fluorescent. Upon cleavage of the probe by incubation with lipoprotein lipase (LPL), fluores-cence can be measured at an excitation of 342 nm and emission of 400 nm.
Lipoprotein lipase (LPL) is the key plasma lipase re-sponsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver. A mutation in the gene coding for LPL can lead to deficiencies in the enzyme, resulting in a diminished ability to breakdown fatty acids. Such LPL deficiency is known as chylomicronemia or Type I hyperlipoproteinemia. Our Lipoprotein Lipase (LPL) ELISA Kit provides a simple, convenient method to measure LPL levels in plasma, serum or other biological fluids from a variety of species (see below). LPL amounts are quantified against the provided LPL Standard in a colorimetric 96-well microplate reader.
Standard Curve Generated with the Lipoprotein Lipase (LPL) ELISA Kit.
Sensitive: Detect as little as 20 ng/mL of LPL Versatile: Suitable for human, rat, bovine, guinea
pig, or chicken samples (but not mouse)
Lipoprotein lipase (LPL) is the key plasma lipase responsible for the hydrolysis of the triglyceride core found in very low density lipoprotein (VLDL) particles formed in the liver.
Lipoprotein Metabolism METABOLISM RESEARCH
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Serum Triglyceride Quantitation Kits
Triglycerides serve as an energy source and play a key role in lipid metabolism. Lipases secreted into the intestines hydrolyze the triglyceride ester bond, pro-ducing glycerol and free fatty acids. Hepatic lipases also break down triglycerides in the liver to assemble very low density lipoprotein (VLDL) particles. Our Serum Triglyceride Quantitation Kits use a cou-pled enzymatic reaction system to measure triglyc-eride concentrations. First, a lipase hydrolyzes the ester bond, yielding free glycerol. The glycerol is then phosphorylated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluorescence-based detection, both of which are per-formed in a 96-well microtiter plate.
Standard Curve Generated with the Serum Triglyceride Quantitation Kit (Colorimetric).
Product Name Detection Size Catalog Number
Colorimetric 100 Assays STA-396
Fluorometric 100 Assays STA-397 Serum Triglyceride Quantification Kit
Sensitive: Detect as little as 10 µM (1 mg/dL) with the colorimetric format and 2 µM (0.2 mg/dL) with the fluorometric format
Versatile: Suitable for serum, plasma, and cell and tissue lysates
Free Fatty Acid (FFA) Assay Kits
Free fatty acids (FFA) are released upon hydrolysis of triglycerides. FFAs then bind plasma albumin for circulation in the body, serving as a readily absorbed energy source for muscle, brain, and other organ tis-sues. Our Free Fatty Acid Assay Kits use a coupled enzy-matic reaction system to measure free fatty acid con-centrations in serum or plasma. Acyl CoA Synthetase catalyzes FFA acylation of CoA. The Acyl-CoA is then oxidized by Acyl CoA Oxidase, producing hydro-gen peroxide, which reacts with the kit’s probe. Kits are available with either colorimetric or fluorescence-based detection, both of which are performed in a 96-well microtiter plate.
Product Name Detection Size Catalog Number
Colorimetric 100 Assays STA-618
Fluorometric 100 Assays STA-619 Free Fatty Acid Assay Kit
Standard Curve Generated with the Free Fatty Acid Assay Kit (Fluorometric).
Want to measure free glycerol content? See our Free Glycerol Assay Kits on page 146.
Recent Product Citations 1. Chellan, B. et al. (2014). IL-22 is induced by S100/calgranulin
and impairs cholesterol efflux in macrophages by downregulat-ing ABCG1. J. Lipid Res. 55:443-454. (STA-396)
2. Marino, A. et al. (2014). ITCH deficiency protects from diet-induced obesity. Diabetes 63:550-561. (STA-396)
METABOLISM RESEARCH Lipoprotein Metabolism
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Acetylcholine Assay Kits
Phosphatidylcholine Assay Kit
Sphingomyelin Assay Kit
Product Name Detection Size Catalog Number
Colorimetric 100 Assays STA-603
Fluorometric 100 Assays STA-602 Acetylcholine Assay Kit
Our Acetylcholine Assay Kits provide a simple, convenient method to quantify acetylcholine in plasma, serum, cell suspensions, or tissue homogenates. Kits are available with either colorimetric or fluorometric detection, both of which are performed in a 96-well microplate reader.
Product Name Detection Size Catalog Number
Phosphatidylcholine Assay Kit Fluorometric 96 Assays STA-600
Phosphatidylcholine is the foremost phospholipid in eukaryotic cell membranes and comprises about 70% of the total phospholipids in plasma lipoproteins. Our Phosphatidylcholine Assay Kit is a simple fluoro-metric assay that measures phosphatidylcholine in plasma, serum, cell suspensions or tissue homoge-nates. Phospholipase D enzyme hydrolyzes phos-phatidylcholine into phosphatidic acid and choline. The choline is then oxidized by choline oxidase to produce hydrogen peroxide, which is detected by a fluorogenic probe in the presence of horseradish per-oxidase (HRP).
Product Name Detection Size Catalog Number
Sphingomyelin Assay Kit Fluorometric 96 Assays STA-601
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Our Sphingomyelin Assay Kit is a simple fluorometric assay that measures sphingomyelin levels in plasma, serum, cell suspensions or tissue homogenates. Sphingomyelinase hydrolyzes sphingomyelin into ceramide and phosphocholine, which in turn is bro-ken down into choline. Choline is enzymatically oxi-dized to produce hydrogen peroxide, which is de-tected with a fluorogenic probe in the presence of horseradish peroxidase (HRP).
Standard Curve Generated with the Sphingomyelin Assay Kit.
Recent Product Citation Winklger, E.A. et al. (2014). Blood-spinal cord barrier disruption contributes to early motor-neuron degeneration in ALS-model mice. PNAS 111:E1035-E1042.
Lipoprotein Metabolism METABOLISM RESEARCH
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Human C-Reactive Protein ELISA Kit
Human Plasminogen ELISA Kit
Product Name Detection Size Catalog Number
Human C-Reactive Protein (CRP) ELISA Kit Colorimetric 96 Assays STA-392
Product Name Detection Size Catalog Number
Human Plasminogen ELISA Kit Colorimetric 96 Assays STA-393
Standard Curve Generated with the Human Plasminogen ELISA Kit.
Standard Curve Generated with the Human C-Reactive Protein (CRP) ELISA Kit.
C-Reactive Protein (CRP) is a serum protein that binds with high affinity to phosphocholine residues as well as other autologous and extrinsic ligands. CRP is a well-established marker of inflammation and tissue damage, and it has been associated with cardiovas-cular disease, atherosclerosis, and other diseases. Our Human C-Reactive Protein (CRP) ELISA Kit pro-vides a simple, convenient method to measure CRP levels in human plasma, serum, or other biological fluids. CRP amounts are quantified against the pro-vided CRP Standard in a colorimetric 96-well mi-croplate reader.
Sensitive: Detect as little as 1 ng/mL of CRP Versatile: Suitable for plasma, serum, or other
biological fluids
Plasminogen is a plasma glycoprotein that plays a role in macrophage recruitment, arterial stenosis, atherosclerosis, aneurysm formation, wound healing, and neovascularization. Plasminogen exists as an inactive proenzyme, but when converted to the active enzyme plasmin it serves to digest fibrin. This activa-tion is catalyzed by tissue-type plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA). Our Human Plasminogen ELISA Kit provides a sim-ple, convenient method to measure plasminogen lev-els in human plasma, serum, or other biological flu-ids. Plasminogen amounts are quantified against the provided Plasminogen Standard in a colorimetric 96-well microplate reader.
Sensitive: Detect as little as 150 pg/mL of plas-minogen
Versatile: Suitable for plasma, serum, or other biological fluids
METABOLISM RESEARCH Lipoprotein Metabolism
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Product Name Size Catalog Number
Human Albumin 100 µg STA-230
Human Albumin, Malondialdehyde Modified 100 µg STA-210
Human C-Reactive Protein 100 µg STA-240
Human Plasminogen 100 µg STA-239
Product Name Detection Size Catalog Number
Goat Anti-Human Albumin Polyclonal Antibody Immunoblot/ELISA 100 µg STA-130
Rabbit Anti-Human C-Reactive Protein Polyclonal Antibody Immunoblot/ELISA 100 µg STA-140
Goat Anti-Human Plasminogen Polyclonal Antibody Immunoblot/ELISA 100 µg STA-139
Human Serum Proteins
Antibodies to Human Serum Proteins
Antibodies are affinity purified.
Human Albumin ELISA Kit
Product Name Detection Size Catalog Number
Human Albumin ELISA Kit Colorimetric 96 Assays STA-383
Standard Curve Generated with the Human Albumin ELISA Kit.
Our human albumin and oxidized albumin were isolated and purified by HPLC. C-reactive protein was isolated from human pleural fluid. Plasminogen was isolated and purified from human plasma following an ultracentrifu-gation procedure.
Human serum albumin (HSA) is the most abundant protein in human plasma, constituting about half the protein in blood serum. It is typically found in concen-trations around 50 mg/mL. Produced in the liver in a preproalbumin state, albumin transports hormones, fatty acids, and other compounds through the circula-tion. It also maintains pH and osmotic pressure. Our Human Albumin ELISA Kit provides a simple, convenient method to measure HSA levels in human plasma, serum, urine, or other biological fluids. Hu-man albumin amounts are quantified against the pro-vided HSA Standard in a colorimetric 96-well mi-croplate reader.
Sensitive: Detect as little as 100 pg/mL of human serum albumin
Versatile: Suitable for plasma, serum, or other biological fluids
Lipoprotein Metabolism METABOLISM RESEARCH
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Lipid Extraction Kit, Chloroform Free
Lipid Quantification Kit
Product Name Size Catalog Number
50 Preps STA-612 Lipid Extraction Kit (Chloroform Free)
Product Name Detection Size Catalog Number
Lipid Quantification Kit Colorimetric 100 Assays STA-613
Traditional methods of extracting lipids from tissues or cells, such as the well-published Folch method, have used chloroform as the extraction solvent. This method has a couple of disadvantages. First, the or-ganic phase ends up below the aqueous phase, cre-ating problematic removal through the upper phase that risks contamination. Second, chloroform has been classified in many places are a probable human carcinogen. Our Lipid Extraction Kit overcomes both disadvan-tages of the Folch method. The kit provides a chloro-form-free extraction method, and the use of proprie-tary organic solvents places the organic phase above the aqueous phase, allowing easy removal without disturbing the aqueous layer. Each kit provides suffi-cient reagents for 50 extractions from 100 µL sample sizes, but reagents may be scaled up for larger sam-ples.
Total Cholesterol Assay Performed on Extracted Lipids. Lipids extracted from fetal bovine serum (FBS) and HEK293 cells were prepared using the traditional Folch method (blue) and the Lipid Extraction Kit (red). Samples were tested for the presence of cho-lesterol in the Total Cholesterol Assay Kit (Cat. #STA-390).
Standard Curve Generated with the Total Lipid Quantification Kit.
Our Lipid Quantification Kit provides a convenient plate-based method to measure the total unsaturated lipid content found in various samples.* The assay uses a sulfo-phospho-vanillin method in which sam-ples are acidified and heated to solubilize and prime the lipids. The lipids then react with vanillin in acidic conditions to form a colorimetric product detectable in a standard 96-well microplate reader. This kit is compatible with plasma and serum sam-ples, or with crude or purified lipids extracted from cells. Each kit provides sufficient reagents for 100 assays including standards and unknown samples.
*Saturated lipids are not detected with this assay.
144 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
METABOLISM RESEARCH Renal Function Assays
Renal Function Assays
Our Renal Function Assays provide a simple, sensitive method to test for various markers related to kidney function:
Uric Acid / Uricase Urea
Creatinine Protein Carbamylation
Uric Acid/Uricase Assay Kit
Uric acid is the final oxidation end product of purine nucleotide metabolism. Uric acid is a potent antioxi-dant that is released during hypoxic conditions and is usually excreted in the urine via glomerular filtra-tion. In the urine, the enzyme uricase metabolizes uric acid to allantoin which is excreted from the body. Our Uric Acid / Uricase Assay Kit is a simple 96-well microplate-based assay for measuring concentra-tions of either uric acid or uricase in serum, plasma or urine samples. Detection is performed in a fluo-rescence-based plate reader.
Product Name Detection Size Catalog Number
Uric Acid/Uricase Assay Kit Fluorometric 400 Assays STA-375
Assay Principle for the Uric Acid / Uricase Assay Kit.
Product Name Detection Size Catalog Number
Urea Assay Kit Fluorometric 192 Assays STA-382
Sensitive: Detect as little as 0.5 µM of uric acid or 1 mU/mL of uricase
Quantitative: Kit includes both uric acid and uricase standards
Urea is the end product of protein nitrogen metabo-lism and is the primary vehicle for removing toxic ammonia from the body. Urea quantitation is one of the most widely applied tests for kidney function evaluation. Our Urea Assay Kit is a simple 96-well microplate-based assay for measuring urea concentrations in serum, plasma, lysates, or urine samples. Detection is performed in standard colorimetric plate reader.
Urea Assay Kit
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1 to 2 1 to 4 1 to 8
OD
63
0 n
m
Plasma and Serum Dilutions
Plasma
Serum
Human Plasma and Serum Samples Tested with the Urea Assay Kit.
Sensitive: Detect as little as 1 mg/dL of urea Quantitative: Measure unknown samples
against a known urea standard curve
145 www.cellbiolabs.com [email protected]
Renal Function Assays METABOLISM RESEARCH
Urinary Creatinine Assay Kit
Product Name Detection Size Catalog Number
Urinary Creatinine Assay Kit Colorimetric 192 Assays STA-378
Creatinine is a metabolite formed from creatine and phosphocreatine (p-creatine). Subsequently creatinine enters the blood and is then excreted by the kidneys via glomerular filtration. Intra-individual variation of creatinine levels is <15% daily, making it a useful marker for normalizing levels of other mole-cules found in the urine. Our Urinary Creatinine Assay Kit is based on the Jaffe reaction between creatinine and alkaline picrate, which produces an orange-red color com-plex that can be easily read by a standard mi-croplate reader. A creatinine standard is provided to allow quantitative measurements of creatinine levels in urine samples. The assay is simple and takes less than one hour to perform.
Creatinine Levels in Urine Samples in the Presence and Absence of Glucose.
Protein Carbamylation ELISA Kit and Antibodies
Product Name Detection Size Catalog Number
OxiSelect™ Protein Carbamylation Sandwich ELISA Colorimetric 96 Assays STA-877
Goat Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody Immunoblot/ELISA 50 µg STA-077
Rabbit Anti-Carbamyl-Lysine (CBL) Polyclonal Antibody Immunoblot/ELISA 50 µg STA-078
Carbamyl Lysine-BSA N/A 10 µg STA-379
Carbamylation is a post-translational modification which occurs throughout the lifespan of proteins in vivo. Carbamylation results from the binding of iso-cyanic acid, which spontaneously arises from high concentrations of urea, to lysine residues of proteins as carbamyl-lysine (CBL). Our Protein Carbamylation Sandwich ELISA Kit is a convenient microplate-based method for the evalua-tion of protein carbamylation in a variety of sample types. In addition, polyclonal antibodies are available for use in Western blot and ELISA applications.
Standard Curve Generated with the OxiSelect™ Protein Carbamylation Sandwich ELISA Kit.
Formation of Carbamyl-Lysine (CBL) During Carbamylation of Proteins.
METABOLISM RESEARCH Alcohol Assays
146 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Alcohol Assay Kits
Our Alcohol Assay Kits provide a convenient plate-based method to measure primary alcohols, including ethanol, in plasma, serum or saliva samples.* The kit uses an enzymatic oxidation reaction that produces hydrogen peroxide, which reacts with the provided probe. Kits are available with either colorimetric or fluores-cence-based detection, both of which are performed in a 96-well microtiter plate. The colorimetric assay can detect alcohol levels as low as 30 µM, while the fluorometric assay detects as low as 15 µM.
Product Name Detection Size Catalog Number
Colorimetric 100 Assays STA-620
Fluorometric 100 Assays STA-621 Alcohol Assay Kit
Free Glycerol Assay Kits
Product Name Detection Size Catalog Number
Colorimetric 100 Assays STA-398
Fluorometric 100 Assays STA-399 Free Glycerol Assay Kit
Standard Curve Generated with the Free Glycerol Assay Kit.
Glycerol is the backbone of triglycerides. Lipases se-creted in the intestines hydrolyze the triglyceride ester bond, producing glycerol and free fatty acids. Our Free Glycerol Assay Kits use a coupled enzy-matic reaction system to measure free, endogenous glycerol concentrations. The glycerol is phosphory-lated and oxidized, producing hydrogen peroxide, which reacts with the probe provided with each kit. Kits are available with either colorimetric or fluores-cence-based detection, both of which are performed in a 96-well microtiter plate.
Standard Curve Generated with the Alcohol Assay Kit. *These assays are not suitable for urine samples.
148 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
PATHOGEN AND TOXIN ASSAYS Viral Core Antigens
Core antigens provide a convenient way to detect and quantify certain infectious viruses. We offer a variety of ELISA kits to measure the core antigen levels in plasma, serum, or purified virus preps. Note: Assays are for research use only, not for clinical diagnostic use.
Virus Core Antigen Detection
QuickTiter™ HIV-1 p24 ELISA Kit
A popular method of quantifying HIV-1 is the p24 ELISA. Our p24 ELISA kits provide a quick, convenient way to quantify the concentration of your lentivirus.
Product Name Detection Size Catalog Number
QuickTiter™ HIV Lentivirus Quantitation Kit (HIV-1 p24 ELISA) Colorimetric 96 Assays VPK-108-H
5 x 96 Assays VPK-108-H-5
QuickTiter™ MuLV Core Antigen ELISA Kit
Product Name Detection Size Catalog Number
QuickTiter™ MuLV Core Antigen ELISA Kit (MuLV p30) Colorimetric 96 Assays VPK-156
Sensitive: Detect as little as 300 pg/mL Fully quantitative: Recombinant core antigen in-
cluded as positive control
Murine leukemia virus (MuLV) is a retrovirus capable of causing cancer in mice and related vertebrates. Recently discovered in humans, xenotropic murine leukemia virus-related virus (XMRV) is closely related to MuLV; the p30 core antigens share 96% identity. Our QuickTiter™ MuLV Core Antigen ELISA Kit spe-cifically measures the level of the p30 core antigen in blood or other samples.
MuLV Core Antigen HIV-1 p24 Core Protein
Hepatitis B Core Antigen Hepatitis C Core Antigen
Standard Curve Generated with the QuickTiter HBV Core Antigen ELISA Kit.
0
0.5
1
1.5
2
2.5
3
0 5 10 15 20
MuL V p30 (ng /mL )
OD 450nm
149 www.cellbiolabs.com [email protected]
QuickTiter™ HBV Core Antigen ELISA Kit
The QuickTiter™ HBV Core Antigen ELISA Kit spe-cifically quantifies the core protein of Hepatitis B vi-rus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.
Sensitive: Detect as little as 1 ng/mL Fast: Results in about 5-6 hours Fully Quantitative: Recombinant core antigen
included as positive control
Product Name Detection Size Catalog Number
QuickTiter™ HBV Core Antigen ELISA Kit 96 Assays VPK-150
5 x 96 Assays VPK-150-5 Colorimetric
QuickTiter HBV Core Antigen ELISA Kit Standard Curve.
Viral Core Antigens PATHOGEN AND TOXIN ASSAYS
QuickTiter HCV Core Antigen ELISA Kit Standard Curve.
Product Name Detection Size Catalog Number
QuickTiter™ HCV Core Antigen ELISA Kit Colorimetric 96 Assays VPK-151
QuickTiter™ HCV Core Antigen ELISA Kit
Sensitive: Detect as little as 1 ng/mL Fast: Results in about 5-6 hours Fully Quantitative: Recombinant core antigen
included as positive control
The QuickTiter™ HBV Core Antigen ELISA Kit spe-cifically quantifies the core protein of Hepatitis B virus. A mouse monoclonal antibody is coated onto an 8 x 12 strip-well plate which allows the flexibility to save some of the wells for later use. The quantity of HBV is compared to the provided HBV Core Antigen Standard.
Recent Product Citation Chiu, C.Y. et al. (2014). Low-dose benzo(a)pyrene and its epox-ide metabolite inhibit myogenic differentiation in human skeletal muscle-derived progenitor cells. Toxicol. Sci. 10.1093/toxsci/kfu003. (STA-357)
Standard Curve Generated with the Aflatoxin Competitive ELISA Kit.
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PATHOGEN AND TOXIN ASSAYS Toxin Assays
OxiSelect™ BPDE Adduct ELISA Kits
Polycyclic aromatic hydrocarbons (PAH) are potent carcinogenic pollutants commonly associated with oil, ciga-rette smoke, and automotive exhaust. Benzo(a)pyrene, the first chemical carcinogen discovered, is converted to benzo(a)pyrene 7,8 diol-9,10 epoxide (BPDE) through a series of enzymatic reactions. BPDE can attack both proteins and DNA, forming adducts that can potentially result in tumor formation.
Product Name Detection Size Catalog Number
OxiSelect™ BPDE Protein Adduct ELISA Kit Colorimetric 96 Assays STA-301
OxiSelect™ BPDE DNA Adduct ELISA Kit Colorimetric 96 Assays STA-357
Standard Curve Generated Using the OxiSelect™ BPDE DNA Adduct ELISA Kit.
Our OxiSelect™ BPDE Adduct ELISA Kits provide a convenient method to measure BPDE adducts with either DNA or proteins from cells or tissues. Quantitation is performed by comparing unknown values against a standard curve generated with the standard provided in each kit.
Our BPDE DNA Adduct ELISA can detect adducts as low as 30 ng/mL.
Our BPDE Protein Adduct ELISA can detect adducts as low as 60 ng/mL.
Aflatoxin ELISA Kits
Aflatoxins are among the most potent genotoxic agents known. They can induce chromosomal ab-berations, micronuclei, sister chromatid exchange, unscheduled DNA synthesis, and chromosomal strand breaks. Aflatoxins can form adducts with both DNA and proteins. Our Aflatoxin Competitive ELISA Kits provide a con-venient method for detection of aflatoxin adducts. Our Aflatoxin Competitive ELISA Kit measures
the total of Aflatoxin B1 and Aflatoxin B2 adducts in protein samples.
Our Aflatoxin DNA Adduct Competitive ELISA Kit detects total Aflatoxin B1-DNA adducts, both ring-opened and ring-closed forms.
Product Name Detection Size Catalog Number
Aflatoxin DNA Adduct Competitive ELISA Kit Colorimetric 96 Assays AKR-351
Aflatoxin Competitive ELISA Kit Colorimetric 96 Assays AKR-350
151 www.cellbiolabs.com [email protected]
Product Page 293 Cell Lines
AAV 47 Adenovirus 50 GFP Stable Expression 123 Lentivirus 59 Retrovirus 67
3-Nitrotyrosine Antibodies 85 ELISA Kit 85
4-HNE Antibodies 92 ELISA Kit 92
6-4PP Quantitation Kits 98 8-Iso-Prostaglandin F2
ELISA 92
8-Isoprostane ELISA Kit 92 8-Nitroguanine ELISA Kit 95 8-OHdG ELISA Kit 94 8-OHG ELISA Kit 95 A549/GFP Cell Line 123 AAV (Adeno-Assoc. Virus)
Cell Line 47 Expression Systems 41-45 Expression Vectors 46 Helper Free Systems 41-45 Packaging Systems 45 Premade Control Viruses 46 Purification Kits 47 Quantitation Kit 48 shRNA Expression 41-45 Titer Kit 48 Transduction Reagent 48
Acetylcholine Assays 140 Active Rac-GEF Assay 114 Adenovirus
Cell Line 50 Expression Systems 49, 81 microRNA Expression 81 Premade Recombinant 50-53 Purification Kits 54 Quantitation Kits 55 RCA Assay 56 shRNA Expression 49 Titer Kits 55 Transduction Reagent 56
Adhesion Assays 10-11 Adipogenesis Assay 27 Advanced Glycation End
Products Assay 88-89 Advanced Oxidation Protein
Products Assay 87 Aflatoxin Assays 150 AGE Assays 88-89 Albumin
Antibody 142 ELISA Kit 142 Protein 142
Product Page Apolipoproteins
Antibodies 133 ELISA Kits 132 Proteins 132
Arf1 Activation Assay 112-113
Arf6 Activation Assay 112-113
AUF1 Retroviral Vector 68 Autophagy Expression
Vectors 30
-Actin Antibody 125 -Galactosidase
Recombinant Adenovirus 50 Reporter Assays 123
ß-Tubulin Antibody 125 Bacterial Protein Extraction
Reagents 127
Biochips for Cell Adhesion 10 Blocking Reagent 126 BPDE
DNA Adduct ELISA 150 Protein Adduct ELISA 150
BrdU ELISA Kit 25 BT-549/GFP Cell Line 123 C3 Expression Vector 116 CA9 Recombinant
Adenovirus 50
c-Abl Retroviral Vector 67 cAMP ELISA Kits 119 Cancer Cell Assays
Angiogenesis 29 Anoikis 23 Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Colony Formation 6-9 Soft Agar 6-9 Tumor Cell Isolation Kit 9 Tumor Sensitivity 8
Carbamyl Lysine Antibodies 145 ELISA Kits 145
Carbonyl Assays 86 Carboxyethyl Lysine ELISA 88 Carboxymethyl Lysine
Assays 89
Catalase Activity Assays 106 Cdc42
Activation Assay 112-
Agarose Beads 115 Recombinant Adenovirus 51 Recombinant Protein 118 Retroviral Vector 69
Product Page Alcohol Assays 146 Aldehyde-Induced DNA
Damage Assays 99
Alkaline Phosphatase Assays 38 Angiogenesis
Recombinant Adenovirus 50 Tube Formation Assay 29
Anoikis Assays 23 Antibodies
Albumin 142 Apolipoproteins 133 Beta-Actin 125 Beta-Tubulin 125 Carboxymethyl Lysine (CML) 89 C-Reactive Protein 142 Flag Tag 125 Fluorescent Proteins 124 GAPDH 125 GFP 124 GST Tag 125 HA Tag 125 His Tag 125 HNE (4-Hydroxynonenal) 92 MDA (Malondialdehyde) 91 Methylglyoxal (MG) 89 Myc Tag 125 Nitrotyrosine 85 Plasminogen 142 RFP 124
Antibody Tools Blocking Reagent 126 Purification Kit 125 Western Stripping Solution 126
Antioxidant Assays Catalase Activity Assay 106 Cellular Antioxidant Asssay 109 Glutathione Assay 108 Glutathione Reductase Assay 108 HORAC Assay 110 ORAC Assay 110 Superoxide Dismutase
Activity Assay 107 Total Antioxidant Capacity
Assay 110 AOPP Assay 87 AP Sites Quantitation Kit 96 Apo(a) ELISA 132 ApoAI ELISA 132 ApoAI/ApoB Duplex ELISA 133 ApoAII ELISA 132 ApoB ELISA 132 ApoCI ELISA 132 ApoCII ELISA 132 ApoCIII ELISA 132 ApoE ELISA 132
PRODUCT INDEX
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Product Page Cell Lines (cont’d)
MEF Feeder Cells 35 NIH3T3/GFP 123 OVCA429/GFP 123 OVCAR-5/RFP 123 Plat-A Retroviral Packaging
Cells 64
Plat-E Retroviral Packaging Cells 64
Plat-GP Retroviral Packaging Cells 64
SKOV-3/GFP-Luc 123 SKOV-3/Luc 123 SKOV-3/RFP 123 SNL Feeder Cells 35 T47D/GFP 123
Cell Migration Assays 12-19 Cell Proliferation Assays 24-25 Cell Transformation Assays 6-7 Cell Viability Assay 22 Cellular Antioxidant Assay 109 Cellular Senescence Assays 23 CETP ELISA Kit 136 cGMP ELISA Kits 119 Checkpoint Kinase Assays 121 Chemotaxis Assays 15, 19
Cholesterol Assays 130-131
Cholesteryl Ester Transfer Protein Assay
136
Clonogenic Tumor Cell Isolation Kit
9
CML (Carboxymethyl Lysine) Antibodies 89 Assays 89
CML-LDL ELISA Kit 93, 134
c-Myc Retroviral Vectors 69 Colony Formation Assays
Cell Transformation Assays 6-7 Hematopoietic Colony
Forming Cell Assay 36
Stem Cell Colony Assay 37 Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8
Comet Assay Kits & Slides 97
Core Antigen Assays 148-
CPD Quantitation Kits 98 C-Reactive Protein
Antibody 142 ELISA 141 Protein 142
Cre Recombinant Adenovirus 50 Creatinine Assay 145 CSK Recombinant
Adenovirus 53
Product Page Cyclic AMP ELISA Kits 119 Cyclic GMP ELISA Kits 119 CytoSelect™ Cell-Based
Assays
Anoikis 23 Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Cell Viability 22 Chemotaxis 15, 19 Colony Formation 6-9 Cytotoxicity 22 Haptotaxis 16 Phagocytosis 28 Soft Agar 6-9 Transmigration 17 Tumor Sensitivity 8 Wound Healing 20
Cytoskeleton Regulation
Activation Assays 112-114
Adenoviruses 51 Expression Vectors 116 Retroviral Vectors 69
Cytotoxicity Assay 22 DCC Recombinant
Adenovirus 51
DNA Damage Assays 6-4PP Quantitation 98 8-Nitroguanine ELISA Kit 95 8-OHdG ELISA Kit 94 Aldehyde-Induced Damage 99 AP Sites Quantitation Kit 96 BPDE Adduct ELISA 99 Comet Assays 97 CPD Quantitation 98 Double-Strand Break Assay 96 UV Damage 98
DNA Methylation Assays 100 ECM (Extracellular Matrix)
Assays
Cell Adhesion Assays 10 Cell Invasion Assays 18-19 Tube Formation Assay 29
Endothelial Tube Assay 29 Epitope Tag Antibodies 125 ERK2
Recombinant Adenovirus 52 Retroviral Vector 68
ERK5 Recombinant Adenovirus
52
ES/EC Cells Alkaline Phosphatase Assays 38 Colony Formation Assays 37 Retroviral Expression
Systems 34
Product Page CEA Recombinant
Adenovirus 50
CEL ELISA Kit 88 Cell-Based Assays
Adhesion 10-11 Angiogenesis 29 Anoikis 23 Cell Contraction 29 Cell Viability 22 Chemotaxis 15, 19 Colony Formation 6-9 Cytotoxicity 22 Haptotaxis 16 Invasion 18-19 Migration 12-19 Phagocytosis 28 Proliferation 24-25 Senescence 23 Soft Agar 6-9 Transformation 6-7 Transmigration 17 Tumor Sensitivity 8 Wound Healing 20
Cell Cycle Adenoviruses 51 Anoikis Assay 23 Cell Viability Assay 22 Cytotoxicity Assay 22 Retroviral Vectors 67 Senescence Assays 23
Cell Fractionation Kits 127 Cell Invasion Assays 18-19 Cell Lines
293AAV 47 293AD 50 293LTV 59 293RTV 67 293/CFP 123 293/GFP 123 293/Luc 123 293/YFP 123 293T/GFP-Puro 123 A549/GFP 123 BT-549/GFP 123 ES-2/GFP 123 HeLa/GFP 123 JK1 Feeder Cells 35 MCF-7/GFP 123 MCF-7/Luc 123 MDA-MB-231/GFP 123 MDA-MB-231/GFP-RFP 123 MDA-MB-231/Luc 123 MDA-MB-231/RFP 123 MDA-MB-436/GFP 123 MDA-MB-436/RFP 123 MDA-MB-468/GFP 123
PRODUCT INDEX
153
Product Page ES-2/GFP Cell Line 123 Ethanol Assays 146 Exoenzyme C3 Expression
Vector 116 Extracellular Matrix Kits
Cell Adhesion Assays 10 Cell Invasion Assays 18-19 Tube Formation Assay 29
Feeder Cells 35 FFA Assay Kits 139 Firefly Luciferase Recom-
binant Adenovirus 50 Flag Tag Antibody 125 Free Fatty Acid Assays 139 Free Glycerol Assays 146 Fyn Recombinant Adenovirus 53 Gap Closure Migration
Assays 13 GAPDH Antibody 125 GEF (Guanine Exchange
Factors) Activation Assays 114 Agarose Beads 115
GFP Antibody 124 ELISA Kit 122 Lentiviral Vectors 59 Quantitation Kits 122 Recombinant Adenovirus 50 Recombinant Lentivirus 59 Recombinant Protein 124 Retroviral Vectors 67 Stable Cell Lines 123
GGA3 Agarose Beads 115 Global DNA Methylation
Assays 100 Glutathione Assay 108 Glutathione Reductase Assay 108 Glycerol Assays 146 Glycoaldehyde-BSA 88 GPCR Signaling Products 119 GST
Antibody 125 Inclusion Body Solubilization
and Renaturation Kit 127
GTPase Assay Kits 112-113
HA Tag Antibody 125 Haptotaxis Assays 16 HBV Core Antigen ELISA 149 HCV Core Antigen ELISA 149 HDL
Assay 131 Lipoprotein, Human 131
HEK 293 Cell Lines AAV 47 Adenovirus 50
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Product Page HEK 293 Cell Lines (cont’d)
GFP Stable Expression 123 Lentivirus 59 Retrovirus 67
HeLa/GFP Cell Line 123 Hematopoietic Colony
Forming Cell Assay 36
Hepatitis B Core Antigen ELISA
149
Hepatitis C Core Antigen ELISA
149
HIF-1
Assays 26, 101
Recombinant Adenovirus 50 High Density Lipoprotein
Assay 131 Protein 131
His Tag Antibody 125 Protein ELISA 125
HIV-1 p24 ELISA Kits 60-61,
148 HNE
Antibodies 92 Assays 92
HNE-LDL Assay 93, 134
hnRNPA0 Retroviral Vector 68 HORAC Assay Kit 110 H-Ras
Activation Assay 112-113
Recombinant Protein 118 HuB Retroviral Vector 68 HuC Retroviral Vector 68 HuD Retroviral Vector 68 HuR Retroviral Vector 68 Hydrogen Peroxide Assays 104 Hydroxyl Radical Antioxi-
dant Capacity Assay 110
Hypoxia Assays 26,
IFN Recombinant Adenovirus 52 IB Recombinant Adenovirus 53 IKK Recombinant Adenovirus 53 IL-2 Recombinant Adenovirus 52 Immunoblot Blocking
Reagent 126
In Vitro Angiogenesis Assay 29 In Vitro Tumor Sensitivity
Assay 8
Inclusion Body Solubilization 127 Induced Pluripotent Stem
Cells
Lentiviral Vectors 33 Retroviral Packaging Cells 32 Retroviral Vectors 32-33
Product Page Invasion Assays 18-19 iPS Cell Reprogramming
Lentiviral Vectors 33 Retroviral Packaging Cells 32 Retroviral Vectors 32-33
JK1 Feeder Cells 35 JNK1
Recombinant Adenovirus 52 Retroviral Vector 68
Klf4 Retroviral Vectors 69 KOSM Viral Vectors 33 K-Ras
Activation Assay 112-113
Recombinant Protein 118 LC3 Expression Vectors 30 LCAT Assays 137 LDH Cytotoxicity Assay 22 LDL
Assays 131 Lipoprotein, Human 131 Oxidized 134
LDL Receptor Assay 135 Lecithin Cholesterol Acyl-
transferase Assays 137
Lentivirus Cell Line 59 Concentration & Purification
Kits 62
Control Plasmids 59 Expression Systems 57-58 Expression Vectors 59 Packaging Systems 58 Premade Control Viruses 59 Purification Kits 62 Quantitation Kits 60-61 shRNA Expression 57-58 Titer Kits 60-61 Transduction Kits 63
Leukocyte Assays Adhesion 11 Transmigration 17
Lin-28 Retroviral Vectors 69 Lipid Extraction Kit 143 Lipid Peroxidation Assays
8-Isoprostane ELISA 92 HNE Adduct ELISA 92 Malondialdehyde (MDA)
Assays 90-91
TBARS Assay 90 Lipid Quantification Assay 143 Lipoprotein Lipase Assays 138 Lipoproteins
Assays 130-133
Human 131-132
Oxidized 134
PRODUCT INDEX
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Product Page ORAC Assay Kit 110 OVCA429/GFP Cell Line 123 OVCAR-5/RFP Cell Line 123 Oxidized LDL
Assay Kits 93, 134
Lipoprotein, Human 131 Oxidized Proteins 89 OxiSelect™ Oxidative
Stress / Damage Assays
Antioxidant Assays 106-110
DNA / RNA Damage Kits 94-100
Lipid Peroxidation Assays 90-93 Protein Oxidation Assays 85-89
ROS Assays 102-
OxLDL Assay 93,
OxPL Assay 134 Oxygen Radical Antioxidant
Capacity (ORAC) Assay 110
p24 ELISA Kits 60-61,
148 p38
Recombinant Adenovirus 52 Retroviral Vectors 68
p53 Recombinant Adenovirus 51 Retroviral Vectors 67
p68 RNA Helicase Adenovirus 51 PABP Retroviral Vector 68 PAK1
PBD Agarose Beads 115 Recombinant Adenovirus 51
PCNA ELISA Kit 25 PCSK9 ELISA Kit 136 Peroxide Detection Assays 104 Phagocytosis Assays 28 Phosphatidylcholine Assay 140 Phospho Antibody Stripping
Solution 126
PhosphoBLOCKER™ Western Blot Blocking Reagent
126
Phosphoproteins Antibody Stripping Solution 126 Blocking Reagent 126 Purification Kit 126
PI3K Retroviral Vector 68 PKC Recombinant
Adenovirus 53
Plasminogen Antibody 142 ELISA Kit 141 Protein, Human 142
Product Page LOX-1 Assay 135 LPL Assays 138 Luciferase
Recombinant Adenovirus 50 Reporter Cell Lines 123
Malondialdehyde Antibodies 91 Assays 90-91
MAP Kinase Signaling Recombinant Adenovirus 52 Retroviral Vectors 68
MAPKAPK2 Recombinant Adenovirus 52 Retroviral Vector 68
MCF-7/GFP Cell Line 123 MCF-7/Luc Cell Line 123 MDA (Malondialdehyde)
Antibodies 91 Assays 90-91
MDA-LDL Assay 93, 134
MDA-MB-231/GFP Cell Line 123 MDA-MB-231/Luc Cell Line 123 MDA-MB-231/RFP Cell Line 123 MDA-MB-436/GFP Cell Line 123 MDA-MB-436/RFP Cell Line 123 MDA-MB-468/GFP Cell Line 123 MEF Feeder Cells 35 MEK1
Recombinant Adenovirus 52 Retroviral Vector 68
MEK5 Recombinant 52
MEKK1 Recombinant Adenovirus
52
MEKK3 Recombinant Adenovirus
52
Methylglyoxal (MG) Antibody 89 ELISA Kits 89
Microfluidic Biochips 10 microRNA Analysis
Adenoviral Expression System 81
Clone Collection 74-80 Control Vectors 80 Expression Vectors 74-80 Functional Reporter System 82 Precursor Clone Collection 74-80 Retroviral Expression Vector 81 Transduction Enhancer 82
Migration Assays 12-19 miRNA Analysis
Adenoviral Expression System 81
Clone Collection 74-80 Control Vectors 80
Product Page miRNA Analysis (cont’d)
Expression Vectors 74-80 Functional Reporter System 82 Precursor Clone Collection 74-80 Retroviral Expression Vector 81 Transduction Enhancer 82
MKK3 Recombinant Adenovirus 52 Retroviral Vector 68
MKK4 Recombinant Adenovirus
52
MKK6 Recombinant Adenovirus 52 Retroviral Vector 68
MKK7 Recombinant Adenovirus
52
MTT Cell Proliferation Assay 24 MuLV p30 Core Antigen
ELISA 148
Myc Tag Antibody 125 MyoD Recombinant
Adenovirus 51
Myogenin Recombinant Adenovirus
51
myr-Akt Retroviral Vectors 68 myr-Rac1
Recombinant Adenovirus 52 Retroviral Vectors 69
NANOG Retroviral Vectors 69 N-Carboxyethyl Lysine
ELISA 88
N-Carboxymethyl Lysine Antibody 89 Assay Kits 89
NFB Recombinant Adenoviruses
53
NIH3T3/GFP Cell Line 123 Nitrated LDL 131 Nitric Oxide Assays 105 Nitroguanine ELISA Kit 95 Nitrotyrosine
Antibodies 85 Assay Kits 85
NOD2 Recombinant Adenovirus
53
N-Ras
Activation Assay 112-113
Recombinant Protein 118 Nuclear / Cytosolic Cell
Fractionation Kits 127
NY-ESO-1 Recombinant Adenovirus
50
Oct-3/4 Retroviral Vectors 69 OHdG ELISA Kit 94 OHG ELISA Kit 95
PRODUCT INDEX
Product Page Plat-A Retroviral Packaging
Cells 64 Plat-E Retroviral Packaging
Cells 64 Plat-GP Retroviral Packaging
Cells 64 Platinum Retroviral
Expression Expression Systems 65 Packaging Cell Lines 64
pMX Retroviral Vectors 66-69 PRAK
Recombinant Adenovirus 52 Retroviral Vector 68
Proliferating Cell Nuclear Antigen Assay 25
Proliferation Assays 24-25 Protease Retroviral Vectors 69 Protein Extraction Reagents 127 Protein Oxidation Assays
Advanced Glycation End Products (AGE) 88-89
Advanced Oxidation Protein Products (AOPP) 87
BPDE Adduct 87 Carbonyl 86 CEL (Carboxyethyl Lysine) 88 CML (Carboxymethyl Lysine) 89 MG (Methylglyoxal) 89 Nitrotyrosine 85
Protein Phosphorylation Antibody Stripping Solution 126 Blocking Reagent 126 Purification Kit 126
Protein Quantitation Kit 128 Proteins
Albumin 142 Apolipoproteins 132 EGFP 124 GRP-PH Domain 119 Oxidized/Nitrated 131
Purification Kits AAV 47 Adenovirus 56 Antibodies 125 Lentivirus 62 Phosphoproteins 126 Retrovirus 70
QuickTiter™ Viral Titer & Quantitation Kits AAV 48 Adenovirus 55 HBV Core Antigen 149 HCV Core Antigen 149 HIV p24 148 Lentivirus, Recombinant 60-61
Product Page
Reporter Genes Lentiviral Vectors 59
Quantitation Assays 122-123
Recombinant Adenovirus 50 Recombinant Lentivirus 59 Retroviral Vectors 67 Stable Cell Lines 123
Retrovirus Concentration & Purification
Kits 70
Expression Systems 65 Expression Vectors 66 Gene-Specific Vectors 67-69 Packaging Cell Lines 64, 67 Purification Kits 70 Quantitation Kits 71 shRNA Expression 66 Transduction Kits 72
RFP Antibody 124 ELISA Kit 123 Recombinant Protein 124 Stable Cell Lines 123
RGS Recombinant Proteins 118 Rho
Activation Assays 112-113
Agarose Beads 115 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vector 69
RhoA Activation Assay 112-113
RhoB Activation Assay 112-113
RhoC Activation Assay 112-113
Rho Kinase Activity Assays 120 RIPA Buffer 128 RNA Damage ELISA Kit 95 RNAi Enhancer Reagent 82 ROCK Activity Assay Kits 120
ROS Assays 102-105
scAAV Control Vector 46 Expression Systems 42-45 Expression Vector 46
SCGE Assay Kits 97 SEAP Recombinant
Adenovirus 50
Senescence Assays 23 shAkt Recombinant
Adenovirus 53
Renal Function Assays 144-
155 www.cellbiolabs.com [email protected]
Product Page
QuickTiter™ Viral Titer &
MuLV p30 148 Retrovirus, Recombinant 71
Rab Recombinant Proteins 118 Rac
Activation Assays 112-113
Agarose Beads 115 GEF Assay 114 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vectors 69
Radius™ Cell Migration Assays
13
Raf1 Recombinant Adenovirus 52 Retroviral Vectors 68
Ral
Activation Assay 112-113
Agarose Beads 115 Recombinant Proteins 118
Ran
Activation Assay 112-113
Agarose Beads 115 Recombinant Protein 118
Rap
Activation Assays 112-113
Recombinant Proteins 118 RAPAd® Adenoviral
Expression Systems 49
Rapid GST Inclusion Body Solubilization and Renatu-ration Kit
127
Rapid RCA Assay 56 Ras Superfamily
Activation Assays 112-113
Agarose Beads 115 Expression Vectors 116 Recombinant Adenovirus 51 Recombinant Proteins 118 Retroviral Vectors 69
RCA Assay Kit 56 Reactive Oxygen Species
(ROS) Assays 102-105
Recombinant Adenoviruses 50-53 Recombinant Proteins
Fluorescent Proteins 124 GRP-PH Domain 119 Small GTPase 118
Rel B Recombinant Adenovirus
53
PRODUCT INDEX
156 Fax 1-858-271-6514 USA Toll-Free 1-888-CBL-0505 Phone 1-858-271-6500
Product Page shRNA Expression
Adeno-Associated Virus 41-45 Adenovirus 49 Lentivirus 57-59 Retrovirus 66
Single Cell Gel Electro-phoresis Assays
97
SKOV-3/GFP-Luc Cell Line 123 SKOV-3/Luc Cell Line 123 SKOV-3/RFP Cell Line 123 Small GTPase
Activation Assays 112-
Active GEF Assays 114 Agarose Beads 115 Expression Vectors 116 Premade Adenoviruses 51 Retroviral Vectors 69
SNL Feeder Cells 35 SOD Activity Assay Kit 107 Soft Agar Colony Assay Kits
Cell Transformation Assays 6-7
Hematopoietic Colony 36
Stem Cell Colony Formation 37
Tumor Cell Isolation Kit 9 Tumor Sensitivity Assay 8
SOK Recombinant 52
Sox2 Retroviral Vector 69 Sphingomyelin Assay 140 Src Recombinant Adenovirus 53 Stat5 Retroviral Vectors 68 Stem Cell Research
Alkaline Phosphatase Detection Kits 38
Feeder Cells 35
Hematopoietic Colony 36
iPS Cell Reprogramming 32-33 PCR Primers 38
Retroviral Expression 34
Stem Cell Colony Assay 37 Total Protein: ES Cell Line D3 38 Total RNA: ES Cell Line D3 38
Superoxide Dismutase Assay 107 T47D/GFP Cell Line 123 TAC Assay 110
Tac-Rac1 Recombinant Adenovirus
52
TBARS Assay Kit 90 TIA1 Retroviral Vector 68 Tiam1 Assay Kit 114 TIAR Retroviral Vector 68
Product Page
Total Antioxidant Capacity 110
Total Cholesterol Assays 130 Total Protein: ES Cell Line D3 38 Total RNA: ES Cell Line D3 38 Transcription Regulation
Retroviral Vectors 68 Transformation Assays 6-7 Transmigration Assays 17 Triglyceride Assays 139 TTP Retroviral Vector 68 Tube Formation Assay 29 Tumor Antigen Adenoviruses 50 Tumor Cell Assays
Cell Adhesion 10-11 Cell Invasion 18-19 Cell Migration 12-19 Cell Transformation 6-7 Chemosensitivity 8 Soft Agar Colony Formation 6-9 Transmigration 17 Tumor Cell Isolation 9
Tyrosine Kinase Adenoviruses 53
uPA / uPAR Retroviral Vectors 69
Urea Assay 144 Uric Acid / Uricase Asay 144 UV DNA Damage Assays 98 V5 Tag Antibody 125 VEGF Recombinant
Adenovirus 50 Very Low Density Lipoprotein
Assay 131 Lipoprotein, Human 131
ViraBind™ Purification Kits AAV 47 Adenovirus 54 Lentivirus 62 Retrovirus 70
ViraDuctin™ Transduction Kits & Reagents AAV 48 Adenovirus 56 Lentivirus 63 Retrovirus 72
Viral Expression Systems AAV 41-45 Adenovirus 49, 81 Lentivirus 57-58 Retrovirus 65
Viral Packaging Cells AAV 47 Adenovirus 50 Lentivirus 59 Retrovirus 64, 67
Product Page Viral Titer Kits
AAV 48 Adenovirus 55 Lentivirus 60-61 Retrovirus 71
Viral Transduction Reagents AAV 48 Adenovirus 56 Lentivirus 63 Retrovirus 72
ViraSafe™ Lentivirus Expression Systems 57-58
Virus Core Antigen Assays HBVcAg 149 HCVcAg 149 HIV-1 p24 148 MuLV p30 148
Virus Purification Kits AAV 47 Adenovirus 54 Lentivirus 62 Retrovirus 70
Virus Quantitation Kits AAV 48 Adenovirus 55 Lentivirus 60-61 Retrovirus 71
VLDL Assay 131 Lipoprotein, Human 131
VSV-G Retroviral Vector 67 Western Blot Blocking
Reagent 126
Wound Healing Assay 20 WST-1 Cell Proliferation
Reagent 24
PRODUCT INDEX
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