advancing life science to foster a better world...(650 nm red, 488 nm blue) 42 kda. anti-beta-actin...

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ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD

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Page 1: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

A D V A N C I N G L I F E S C I E N C E T O F O S T E R A B E T T E R W O R L D

Page 2: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

Revitablot™ Stripping BufferTest with Secondary HRP-conjugated, Dylight®-conjugated, and IRDye®-conjugated Antibodies

Page 3: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

Revitablot™ Product Specifics

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Fast | Strips most proteins in as little as 10 minutes Efficient | Minimal optimization required Easy | Ambient incubation requires little intervention Safe | Non-toxic materials Multiple Applications | Effective for both Nitrocellulose and PVDF

membranes Multi-Use | Can be used to strip at least four times

Page 4: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

L A N E 1 :

L A N E 2 :

L A N E 3 :

7.5 μg HCT ꟷ 116 cell lysates

22.5 μg HCT ꟷ 116 cell lysates

15 μg HCT ꟷ 116 cell lysates

Colorectal carcinoma HCT-116 whole cell lysates were analyzed on a SDS-PAGE 4ꟷ20% gradient gel (Bio-Rad) and transferred to nitrocellulose membranes. The lysates were followed by blocking with 5% BLOTTO (#B501-0500) for 1 hour at room temperature. The membranes were first probed with Anti-AKT pS473 (MOUSE)(#200-343-268) at a dilution of 1:1,000 overnight at 4°C. Imaging was performed after drying membranes at 650 nm. Membranes were then incubated with Revitablot™ for 5 minutes at room temperature, followed by re-blocking with 5% BLOTTO for 2 hours at room temperature. Secondary probing was performed with Anti-Beta-Actin (RABBIT) (#CUST17) at a dilution of 1:1,000 for 4 hours at room temperature. Membranes were then dried, imaged at 488 nm and 650 nm.

Efficiency in WB

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(650 nm Red, 549 nm Green)60 kDaAnti-AKT pS473 (MAb) DyLight™ 649 Conjugated

(650 nm Red, 488 nm Blue)42 kDaAnti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated

Page 5: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

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1. Image for the primary target and then wash blot for at least 10 minutes in 1x TBS-T on an orbital shaker at room temperature. This process removes any remaining HRP substrate.

2. Decant TBS-T and apply 10 mL of Revitablot ™Note: Volume depends on size of blot; apply enough buffer to sufficiently cover the membrane. Incubate from 5ꟷ10 minutes (user determined) on an orbital shaker at room temperature.

3. Decant Revitablot™ buffer and wash blot for at least 10 minutes in 1X TBS-T as in Step 1.

4. Decant TBS-T and block blot in 5–10 mL of 5% BLOTTO (#B501-0500) for 2 hours on an orbital shaker at room temperature.

HRP-Conjugated & Fluorescent-Conjugated Antibodies Protocol

Page 6: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

HRP-Conjugated & Fluorescent-Conjugated Antibodies Protocol (Cont.)

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5. Decant the BLOTTO and apply the next primary antibody target. Incubate for the optimum time and conditions per provided instructions.

6. Complete the Western blot procedure (washing, application of secondary antibody, substrate addition, and imaging). Note: Blots may be stripped up to four times for HRP-Conjugated antibodies and up to two times for fluorescent-conjugated antibodies.

Page 7: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

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Revitablot™ Competitor

Initial TargetSTAT5

(#600-401-A44)

95 kDa

Revitablot™ Competitor

Re-Probe forα-Tubulin

(#200-301-880)

95 kDaIncubation Blots with

Stripping Buffers(20 minutes)

Revitablot vs. Competitor

L A N E 1 : 70 μg

L A N E 2 : 10 μg

Page 8: ADVANCING LIFE SCIENCE TO FOSTER A BETTER WORLD...(650 nm Red, 488 nm Blue) 42 kDa. Anti-Beta-Actin pS473 (PAb) DyLight™ 488 Conjugated. 5 1. Image for the primary target and then

THANK YOUV I S I T U S O N S O C I A L !

Slide 1Fig. 16.1, p.488. Slide 2Fig. 16.2, p.488

Summer Internship Course Requirements IOE 488/489 CNM 488

ESP-488 Software Reference Manual for LynxOS and the AT-GPIB · 2018. 10. 18. · ESP-488 ESP-488 is used throughout this manual to refer to ESP-488 for LynxOS. IEEE 488 and IEEE

GOOD BAD - Cornell Center for Materials Research · To move the wheel to the GOOD position, first choose the laser “785 nm Linefocus” if you are using 785 nm or “SM 488 nm Raman

Boulevard du Temple - Start - BioVis · Confocal Illumination Lasers monochromatic coherent types: gas, solid, diode Argon Ion 353-361, 488, 514 nm Krypton -Argon 488, 568, 647 nm

T E District 6 Feet - City and County of Denver Official …€¦ · XW XW XW XW XW XW XW nmn nm nm nm nm nm nm nm n nm nm nmn n nm nm nm nm nm nm nm nm nm nm n nm nm nm nm nm n nm

Ward / Quartier 1...$K $K $K $ $K!C nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm …

Amplification by - gazigoz.org · Ankara, Mart 2010 400 nm Excimer 193 nm 700 nm 100 nm Argon 488-514 nm Diode lazer 810 nm Nd:YAG 1064 nm(NIR) CO2 10.600 nm (FIR)

USD 259 District Map...nn n n n n n n n n n n n n n n n n n n nm nmnm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nmn nm nmnm nm nm nmnnm

Renishaw inVia - Welcome to the University at …...2.0 Specifications/Features Three laser wavelengths: 488 nm, 514 nm, 785 nm Raman shift range: up to 4000 cm-1 Photoluminescence

MAPA DE VALORES DE TERRENOS POR ZONAS … del Canton Alajuela 0… · nm nm æ nm nm æ nm nm nm nm æ æ nm nm nm nm nm æ nm nm nm nm æ nm nm nm nm æ æ æ æ nm nm æ nm nm nm

11°30'0W 11°0'0W K en m aD is tr c M p 3 5 . 7 kk Th is m ... · nm nm nm nm nm nm nm nm nm nm nm nm nm nm nmnm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm

EFFECTS OF DIFFERENT LIGHT SPECTRA ON THE GROWTH, … · 2016-06-29 · In this study, effects of three different light spectra: white (400-700 nm), blue (407-488 nm), and red (621-700

BMC Medical Imaging BioMed Central...detector array 1344 × 1024 px (Hammamatsu, Japan), a 3-line Argon-Krypton Laser; 488 nm, 568 nm, 647 nm (Melles Griot, USA) and a mercury lamp

Boundary-breaking acoustic focusing cytometry · 2021. 2. 25. · 710/50 Qdot® 705 Blue 488 nm 488/10 FSC 488/10 SSC*** 530/30 FITC EGFP, Emerald GFP 574/26* Propidium iodide EYFP

DyLight Immunofluorescent Detection - Biocare Medical

Sponsored and reviewed by ICCS Quality and Standards ... (488 nm) with emission maxima of 617 nm (PI) and 647 nm (7-AAD).8-10 Gating on viable events is performed by drawing a gate

MAPA DE VALORES DE TERRENOS Y … san jose goicoechea... · nm æ æ nm nm æ nm nm nm æ nm æ nm nm nm æ nm nm nm nm æ nm æ nm nm nm æ nm nm æ æ æ æ æ nm nm æ æ æ nm

Serial specification of diverse neuroblast identities from a … · conjugated to Dylight 488, 549, 594 and 750 (Pierce) and Alexa Fluor 647 fluorescent conjugates (Invitrogen) diluted

Systems for industry & research - GMP SA GMP.pdfModu-Laser – Air-cooled compact Argon & Argon/Krypton lasers. Selec-table multiline operation, 458 nm, 488 nm, 514 nm, 568 nm, 647nm

Senior Consultant Refractive Surgery Service Singapore ... · Senior Consultant Refractive Surgery Service ... 1064 nm Argon 488-514 nm Green HeNe 543 nm CO 2 10,600 nm Diode 680

3403000 3403500 3404000 3404500 3405000 3405500 0 5 · 2017. 10. 17. · nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nmnm nm nm nmnm nmnm nm nm nm nm nm nm nmnm

Supporting Information The effects of protein charge …...Fluorescence of GFP mutants (λ ex = 488 nm, λ em = 530 nm, Gain = 50) was used to measure relative protein concentration.4

Carbohydrate Labeling and Analysis Kit - sciex.com · Pharmaceutical Analysis System, using the 488 nm laser and the 520 nm emission filter. This kit contains the reagents, buffer,

labeling and detection - Fisher Scientific...glowing advantages Table 1. Spectral properties of Thermo Scientific DyLight Fluorescent Dyes. Emission DyLight Dye Ex/Em* ε† Spectrally

Spectralis Hardware Operating Manual · 1. For excitation of fluorescein, a blue solid state laser (wavelength 488 nm) is used. A barrier filter at 500 nm separates excitation and

geospasial.bnpb.go.idgeospasial.bnpb.go.id/.../PETA-SEBARAN-PENDIDIKAN-DI-SEKITAR-GUNUNG... · nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm nm

High-fidelity spectroscopy at the highest resolutionsdainis/Presentations/Potsdam_2009_Spectroscopy.pdf · Granulation near the limb (towards the top) at 488 nm; Swedish 1-m solar

guava Flow Cytometry easyCyte Systems - Biotecha.lt...The guava easyCyte family of flow cytometry uses one or two lasers, blue (488 nm) or blue (488 nm) and red (640 nm) to achieve

CYTOSKELETON NEWS · Acti-stain™ 488 phalloidin 480 nm 535 nm 57 PHDG1-A 300 Slides Acti-stain™ 535 phalloidin (Rhodamine Phalloidin) 535 nm 585 nm 27 PHDR1 300 Slides Acti-stain™

NeurobiologyofDisease HeterogeneousIntegrationofAdult ...zoom. Z-series stacks were captured at 0.2 m increments. Two-channel sequential scans were captured using 488 and 543 nm laser

Snippetz_Issue 488

I 60204 U.S DATA 488~ AND 488~ SHEET SONOBAR Noise

Mapping Neurotransmitter Identity in the Whole-Mount ... · generally as follows: AF488 with a 488-nm laser and 498– 543-nm detection, DL550 and Cy3 with a 561-nm laser and 569–595-nmdetection,CF594witha594-nmlaserand600–