antibodies and elisa

Toll Free Number (in the United States) • (800) 436-9910 • (609) 497-0253 • Fax Orders (609) 497-0321 1

Manufacturer of Quality Cytokine Products

Published by:

PeproTech Inc.

Rocky Hill, New Jersey USA

3rd Edition, © 2012

Antibody Resource Guide

2 Toll Free Number (in the United States) • (800) 436-9910 • (609) 497-0253 • Fax Orders (609) 497-0321

ELISA Buffer Kit ..................................................................... 3

PeproTech Worldwide Directory ............................................ 4

Ordering Information ............................................................. 5

Affinity-Purified Polyclonal/Biotinylated Antibodies ............... 6

Monoclonal Antibodies ...................................................... 23

ELISA Development Kits ...................................................... 25

Mini ELISA Development Kits .............................................. 27

Appendices: ................................................................... 28-63

Appendix I (Antigen Affinity Purification of Antibodies) .............. 28

Appendix II (ELISA) .............................................................. 32

Appendix III (Western Transfer) .............................................. 40

Appendix IV (Immunohistochemistry) ..................................... 49

Appendix V (Microarray) ....................................................... 56

Appendix VI (Flow Cytometry) ............................................... 59

PeproTech Publications ....................................................... 64

Table of Contents

This publication describes the production, application, and

availability of PeproTech’s anti-cytokine antibodies and ELISA

Development Kits. For the complete list of our products, please

visit www.peprotech.com.


The buffers in this supplemental kit have been specially formulated for optimal performance when used in conjunc-tion with PeproTech’s ELISA Development Kits. The buffer kit contains all of the necessary components to assay ten 96-well ELISA plates (included) and contains detailed handling instruc-tions. All of the reagents have been sterile filtered to minimize assay interference and maximize shelf life. The ELISA Buffer Kit can be purchased along with PeproTech’s ELISA Development Kit at a discounted price.

This easy-to-use ELISA buffer kit can also be purchased as a stand-alone product since the plates included are not pre-coat-ed with capture antibody. The flexibility of this format allows the researcher to develop and optimize an assay for use with their own capture and detection antibodies. When using the ELISA

Buffer Kit in this capacity the actual antibody concentrations and detection ranges of the ELISA will vary according to the specific components used.

Kit Contents: - 20X Plate Coating Buffer

- 1X Blocking Buffer

- 20X Sample Diluent

- 20X Wash Buffer

- Ready-to-use ABTS

- 10 Sterile, Uncoated 96 well ELISA Plates

- 50 Plate Sealing Films

- Buffer Handling Instructions

ELISA Buffer Kit.

ELISA Buffer Kit Catalog # 900-K00 PRICE: $195.00

ELISA Buffer Kit and EDK Combination PRICE: $450.00

PeproTech’s - ELISA Buffer Kit


PeproTech Worldwide Offices

NordicKlarabergsviadukten 70 107 24 StockholmSwedenTel: +46 (0)8 640 41 07Fax: +46 (0)8 640 41 09email: [email protected]

USA & JapanPrinceton Business Park5 Crescent Ave.P.O. Box 275Rocky Hill, NJ 08553-0275 U.S.A.Tel: (800) 436-9910 (609) 497-0253Fax: (609) 497-0321email: [email protected]@peprotech.comwww.peprotech.com

Asia, Africa & Australia12 Hamada StreetTamar BuildingRehovot, 76703Israel Tel: +972 8 946 0948Fax: +972 8 946 0861email: [email protected]

Latin & South America Luz Saviñon No. 1027-4Col. NarvarteDel. Benito JuarezMéxico, DFTel: (52) (55) 5672-0389Fax: (52) (55) 5672-0388email: [email protected]

EuropePeproTech House29 Margravine RoadLondon W6 8LLEnglandTel: +44 (0)20 7610 3062Fax: +44 (0)20 7610 3430email: [email protected]

Canada4280, boul. Saint JeanSuite 203Dollard des OrmeauxQuebecCanada H9H 2A5Tel: (877) 696-1982 Fax: (877) 696-6998email: [email protected]

Korea 1017, 3rd ComplexKing’s Garden B/DNaesu-Dong, Jongro-Gu,Seoul 110-872, KoreaTel: 82-2-3210-2808Fax: 82-2-3210-2835e-mail: [email protected] www.peprotech.com

GermanyOberaltenallee 8D-22081 HamburgGermanyTel: +49 (0)40 734 35 77 70 +49 (0)800 436 99 10 Fax: +49 (0)40 734 35 77 79email: [email protected] [email protected]

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ChinaRoom 1918 Gold River Int. BuildingNo. 35, Shishan RoadSuzhou, Jiangsu ProvinceP. R. China, 215011Tel: +86 512 6832 5993Fax: +86 512 6832 5983email: [email protected]

AustriaSchwendergasse 171150 WienAustriaTel: +43 (0)1 405 9696Fax: +43 (0)1 405 9898email: [email protected]

Czech RepublicGemini - Budova BNa Pankráci 129/1724140 00 - Praha 4Czech RepublicTel: +420 225 992 284Fax: +420 225 992 201email: [email protected] www.peprotech.com


Ordering Information

Prices and Payment Terms:Prices will remain in effect until December 31, 2012.All prices listed in this catalog are in U.S. Dollars. Prices are sub-ject to change without notice. Prices in this catalog are not valid for distributors. Payment terms are net 30 days from receipt of invoice.

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Phone: (800) 436-9910, or (609) 497-0253

Fax: (609) 497-0321

Website: www.peprotech.com

E-mail: [email protected]

[email protected]

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1.5% interest per month may be charged on past due accounts.

VISA MasterCard

and American Express are accepted.

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Warranty:PeproTech warrants to the original purchaser that products sold substantially conform to specifications provided with shipped products and are free from material defects. This warranty does not extend to any product which has been altered in anyway by personnel other than PeproTech's employees or to any product which has been handled in a manner contrary to instructions included in product documentation. PeproTech is not responsible for patent infringement or other violations which may occur with the use of these products. PeproTech makes no warranty of the product's suitability for any purchaser's particular use; suitability for use must be determined by the purchaser or user of the products. All other warranties, expressed or implied are disclaimed to the fullest extent allowed by law. Purchaser's sole remedy under the above warranty shall be replacement with a conforming product, a credit toward future purchases of PeproTech products or, at PeproTech's sole discretion, a refund of the price of the product. If there is a question or concern please contact our Technical Support Department immediately so we may assist you.

Resale:In the absence of an expressed written agreement to the contrary, all products are sold by PeproTech Inc. for the exclusive use of the purchaser and are not to be resold.

Information Changes:This catalog contains current information on PeproTech products. PeproTech reserves the right to discontinue sales of any product or to change product descrip-tions and/or formulations at any time without notice to purchasers.

Product Use Limitations:PeproTech products are offered for research use only. Not for drug use or clinical diagnostic procedures.


Polyclonal/Biotinylated Antibodies Antigen Affinity-Purified

SIzE a SIzE BPRoduCt Catalog $195 $250 PRICEdESCRIPtIon numBER PER VIal PER VIal PER 1mg SouRCE aPPlICatIonS

Anti-human AITRL ...................................... 500-P244 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NA Biotinylated Anti-hAITRL ............................ 500-P244bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA Anti-human ApoE3 ..................................... 500-P238 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NA Biotinylated Anti-hApoE3 ........................... 500-P238bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA Anti-human APRIL ...................................... 500-P192 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, I Biotinylated Anti-hAPRIL............................ 500-P192bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA Anti-human Artemin ................................... 500-P245 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NA Biotinylated Anti-hArtemin ......................... 500-P245bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA Anti-human 4-1BBL .................................... 500-P169 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-h4-1BBL .......................... 500-P169bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human 4-1BB Receptor ....................... 500-P167G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-h4-1BB Receptor ............... 500-P167Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine B71 .......................................... 500-P128 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mB71 .............................. 500-P128bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BAFF ...................................... 500-P163G ............. 50 μg ............ 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hBAFF ............................ 500-P163Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human BCA-1 ...................................... 500-P141 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hBCA-1 ........................... 500-P141bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BD-1 ....................................... 500-P253 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hBD-1 ............................ 500-P253bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BD-2 ....................................... 500-P161G ............. 50 μg ............ 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hBD-2 ............................ 500-P161Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human BD-3 ....................................... 500-P241 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hBD-3 ............................ 500-P241bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BD-4 ........................................ 500-P268 ..................50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated anti-hBD-4 .............................. 500-P268bt ...............25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BDNF ...................................... 500-P84 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hBDNF ............................ 500-P84bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Betacellulin ............................. 500-P254 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hBetacellulin ................... 500-P254bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BMP-2 ..................................... 500-P195 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hBMP-2 ........................... 500-P195bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N

eutralization data not available, I = IHC




Anti-human BMP-7/OP-1............................ 500-P198 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hBMP-7/OP-1 ................. 500-P198bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human BRAK ...................................... 500-P237 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hBRAK ........................... 500-P237bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine C-10 ........................................ 500-P112 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mC-10 ............................. 500-P112bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Cardiotrophin-1 ...................... 500-P101 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hCardiotrophin-1 ................ 500-P101bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sCD22 .................................... 500-P227 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hsCD22 .......................... 500 P227bt ............. 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sCD40 Ligand ........................ 500-P142 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hsCD40 Ligand .............. 500 P142bt ............. 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sCD40 Ligand ........................ 500-P142G ............. 50 μg ............ 100 μg ............. $2,100........................goat ..................... E, WB, NA, IBiotinylated Anti-hsCD40 Ligand .............. 500 P142Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human CNTF ....................................... 500-P140 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hCNTF ............................ 500-P140bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat CNTF.............................................. 500-P79 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat CNTF ......................... 500-P79bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human CTACK ..................................... 500-P294 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hCTACK .......................... 500-P294bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human CTGF ....................................... 500-P252 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hCTGF ............................. 500-P252bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human CTGFL/WISP-2 ....................... 500-P212 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hCTGFL/WISP-2 ............. 500-P212bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine CTLA-4 .................................... 500-P147 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mCTLA-4 ......................... 500-P147bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human CXCL16 ................................... 500-P200 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hCXCL16 ......................... 500-P200bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine CXCL16 ................................... 500-P201G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-mCXCL16 ........................ 500-P201Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human sDDL4 ..................................... 500-P279 ................. 50 ug ............. 100 ug ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hsDLL4 ........................... 500-P279bt .............. 25 ug ............... 50 ug ............. $4,000.................... rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human EGF ......................................... 500-P45 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hEGF ............................... 500-P45bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine EGF ........................................ 500-P174G ............. 50 μg ............ 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mEGF ............................. 500-P174Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-rat EGF ................................................ 500-P277 ................. 50 μg ............. 100 μg ............. $2,100.................... rabbit ....................... E, WB, NBiotinylated Anti-rat EGF ........................... 500-P277bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human EGF receptor ........................... 500-P306 ................. 50 ug ............. 100 ug ............. $2,100.................... rabbit ......................E, WB, NABiotinylated Anti-hEGF receptor................. 500-P306bt .............. 25 ug ............... 50 ug ............. $4,000.................... rabbit ......................E, WB, NAAnti-human EG-VEGF ................................ 500-P188G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hEG-VEGF ...................... 500-P188Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human EMAP-II ................................. 500-P172G ............. 50 μg ............ 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hEMAP-II ...................... 500-P172Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human ENA-78 ................................... 500-P91 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hENA-78 ......................... 500-P91bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Endostatin ............................... 500-P262 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hEndostatin .................... 500-P262bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Eotaxin .................................... 500-P41 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hEotaxin ......................... 500-P41bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Eotaxin .................................... 500-P41G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hEotaxin ......................... 500-P41Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine Eotaxin ................................... 500-P67 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mEotaxin ........................ 500-P67bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Eotaxin-2 ................................ 500-P103G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hEotaxin-2 ...................... 500-P103Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine Eotaxin-2 ............................... 500-P175G .............. 50 μg ............ 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mEotaxin-2 .................... 500-P175Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Eotaxin-3 ................................ 500-P156G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hEotaxin-3 ...................... 500-P156Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Exodus-2 ................................. 500-P109 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hExodus-2 ...................... 500-P109bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine Exodus-2 ................................. 500-P114 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mExodus-2 ..................... 500-P114bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human sFasL/Apo-I ............................. 500-P184G .............. 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hsFasL/Apo-I .................. 500-P184Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human sFas Receptor .......................... 500-P295 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hsFas Receptor ................ 500-P295bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human FGF-acidic .............................. 500-P17 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hFGF-acidic .................... 500-P17bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human FGF-basic ................................ 500-P18 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hFGF-basic ..................... 500-P18bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human FGF-4 ...................................... 500-P158 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hFGF-4 ............................ 500-P158bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine FGF-9 ...................................... 500-P66 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mFGF-9 ........................... 500-P66bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human FGF-10 ................................... 500-P151G ............. 50 μg ............ 100 μg ............. $2,100........................goat ..................... E, WB, NA, IBiotinylated Anti-hFGF-10 ........................ 500-P151Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human FGF-16 ................................... 500-P160G ............. 50 μg ............ 100 μg ............. $2,100........................goat .......................... WB, NBiotinylated Anti-hFGF-16 ........................ 500-P160Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat ......................... WB, NAAnti-human FGF-17 ................................... 500-P152 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hFGF-17 ........................ 500-P152bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human FGF-17 ................................... 500-P152G ............. 50 μg ............ 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hFGF-17 ........................ 500-P152Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human flt3-Ligand .............................. 500-P42 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hflt3-Ligand .................... 500-P42bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Follistatin ................................ 500-P207 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hFollistatin ...................... 500-P207bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Fractalkine .............................. 500-P98 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hFractalkine .................... 500-P98bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human gAcrp30/Adipolean ................. 500-P193G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hgAcrp30/Adipolean ............... 500-P193Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Adipolean Variant ................... 500-P205 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hAdipolean Variant ................ 500-P205bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Galectin-1 ............................... 500-P210 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hGalectin-1 ..................... 500-P210bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human Galectin-3 ............................... 500-P246 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hGalectin-3 ..................... 500-P246bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GCP-2...................................... 500-P120 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hGCP-2 ........................... 500-P120bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human G-CSF ...................................... 500-P43 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hG-CSF ........................... 500-P43bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine G-CSF ..................................... 500-P69 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mG-CSF .......................... 500-P69bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GDF-3 ..................................... 500-P235 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hGDF-3 ........................... 500-P235bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GDNF ...................................... 500-P81 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hGDNF ........................... 500-P81bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GM-CSF .................................. 500-P33 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hGM-CSF ........................ 500-P33bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine GM-CSF .................................. 500-P65 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mGM-CSF ....................... 500-P65bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat GM-CSF ......................................... 500-P225 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rGM-CSF ......................... 500-P225bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GRO/MGSA ............................. 500-P92 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hGRO/MGSA .................. 500-P92bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat GRO/KC ......................................... 500-P74 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat GRO/KC ..................... 500-P74bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GROβ ..................................... 500-P104 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hGROβ ........................... 500-P104bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat GROβ/MIP-2 .................................. 500-P75 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat GROβ/MIP-2 ................. 500-P75bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human GROγ ...................................... 500-P105 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hGROγ ........................... 500-P105bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human HCC-1 ..................................... 500-P106 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hHCC-1 ........................... 500-P106bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Heregulinβ-1 ........................... 500-P288 ................. 50 μg ............. 100 μg ............. $2,100.................... rabbit ..................... E, WB, N, IBiotinylated Anti-hHeregulinβ-1 ................. 500-P288bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human I-309 ....................................... 500-P110 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hI-309 ............................. 500-P110bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human ICAM-1 ................................. 500-P287 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hICAM-1 ....................... 500-P287bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IFN-β .................................... 500-P32B ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIFN-β .......................... 500-P32Bbt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IFN-γ ..................................... 500-P32 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIFN-γ .......................... 500-P32bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IFN-γ ..................................... 500-P119 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIFN-γ ......................... 500-P119bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IFN-γ ............................................. 500-P122 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-rat IFN-γ ........................ 500-P122bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IFN-γ ............................................. 500-P122G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-rat IFN-γ ........................ 500-P122Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IFN-λ2 .................................... 500-P247 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hIFN-λ2 ......................... 500-P247bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IGF-I ....................................... 500-P11 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIGF-I ............................ 500-P11bt ............... 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IGF-I ....................................... 500-P157G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mIGF-I ............................ 500-P157Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IGF-II ...................................... 500-P12 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIGF-II ............................ 500-P12bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IGF-BP1 .................................. 500-P228 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIGF-BP1 ........................ 500-P228bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IGF-BP3 .................................. 500-P230 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hIGF-BP3 ....................... 500-P230bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IGF-BP5 .................................. 500-P232 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIGF-BP5 ....................... 500-P232bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IGF-BP7 .................................. 500-P234 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIGF-BP7 ....................... 500-P234bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-1α ..................................... 500-P21A................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-1α ........................... 500-P21Abt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-murine IL-1α ..................................... 500-P51A................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-1α .......................... 500-P51Abt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-1α ............................................ 500-P180G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-rat IL-1α ........................ 500-P180Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine IL-1β ..................................... 500-P51 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-1β .......................... 500-P51bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-1β ............................................ 500-P80 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat IL-1β ....................... 500-P80bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-1RA .................................. 500-P209 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-1RA ........................ 500-P209bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-2 ......................................... 500-P22 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-2 ............................... 500-P22bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-2 ......................................... 500-P22G ................ 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hIL-2 ............................... 500-P22Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine IL-2 ......................................... 500-P111 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-2 .............................. 500-P111bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-2 ................................................ 500-P274 ................. 50 μg ............. 100 μg ............. $2,100.................... rabbit ....................... E, WB, NBiotinylated Anti-rat IL-2 ............................ 500-P274bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human IL-3 ......................................... 500-P23 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-3 ............................... 500-P23bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-3 ......................................... 500-P53 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-3 .............................. 500-P53bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-3β ............................................. 500-P177G ............. 50 μg ............ 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-rat IL-3β ......................... 500 P177Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IL-4 ......................................... 500-P24 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-4 ............................... 500-P24bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-4 ......................................... 500-P54 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-4 .............................. 500-P54bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-4 ................................................ 500-P94 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-rat IL-4 ............................ 500-P94bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-5 ......................................... 500-P25 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-5 ............................... 500-P25bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-murine IL-5 ......................................... 500-P55 ................... 50 ug ............. 100 ug ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-5 .............................. 500-P55bt ................ 25 ug ............... 50 ug ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-6 ......................................... 500-P26 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-6 ............................... 500-P26bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-6 ......................................... 500-P26G ................ 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hIL-6 ............................... 500-P26Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine IL-6 ......................................... 500-P56 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-6 .............................. 500-P56bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-6 ................................................ 500-P73 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat IL-6 ............................ 500-P73bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-6 ................................................ 500-P73G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-rat IL-6 ............................ 500-P73Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IL-7 ......................................... 500-P27 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-7 ............................... 500-P27bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-7 ......................................... 500-P57 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-7 .............................. 500-P57bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-8 ......................................... 500-P28 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-8 ............................... 500-P28bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-9 ......................................... 500-P29 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-9 ............................... 500-P29bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-9 ......................................... 500-P59 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-9 .............................. 500-P59bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-10 ....................................... 500-P20 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-10 ............................. 500-P20bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-10 ....................................... 500-P60 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-10 ............................ 500-P60bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IL-10 .............................................. 500-P139 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-rat IL-10 .......................... 500-P139bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-11 ....................................... 500-P01 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-11 ............................. 500-P01bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-12 ....................................... 500-P154G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hIL-12 ............................. 500-P154Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-murine IL-12 ....................................... 500-P155G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mIL-12 ............................ 500-P155Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IL-13 ....................................... 500-P13 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-13 ............................. 500-P13bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-13 ...................................... 500-P178 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-13 ........................... 500-P178bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-15 ....................................... 500-P15 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-15 ............................. 500-P15bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-15 ....................................... 500-P173 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-15 ............................ 500-P173bt ............ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-16 ....................................... 500-P06 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-16 ............................. 500-P06bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17A ..................................... 500-P07 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-17A ........................... 500-P07bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17A ..................................... 500-P07G ................ 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hIL-17A ........................... 500-P07Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine IL-17A ..................................... 500-P265 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mIL-17A .......................... 500-P265bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17B ..................................... 500-P248 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIL-17B ........................... 500-P248bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17D..................................... 500-P88 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIL-17D .......................... 500-P88bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17E ..................................... 500-P89 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIL-17E ........................... 500-P89bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-17F ..................................... 500-P90 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-17F ........................... 500-P90bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-18BP ................................... 500-P153G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-mIL-18BP ........................ 500-P153Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human IL-19 ....................................... 500-P189 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIL-19 ............................. 500-P189bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-20 ....................................... 500-P190G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hIL-20 ............................. 500-P190Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human IL-21 ....................................... 500-P191 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-21 ............................. 500-P191bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-Murine IL-21 ....................................... 500-P278 ................. 50 μg ............. 100 μg ............. $2,100.................... rabbit ......................E, WB, NABiotinylated Anti-mIL-21 ............................ 500-P278bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human IL-22 ....................................... 500-P211 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hIL-22 ............................. 500-P211bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IL-22 ....................................... 500-P223 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIL-22 ............................ 500-P223bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-31 ....................................... 500-P249 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hIL-31 ............................. 500-P249bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IL-33 ....................................... 500-P261 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIL-33 ............................. 500-P261bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human IP-10 ....................................... 500-P93 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hIP-10 ............................. 500-P93bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine IP-10 ....................................... 500-P129 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mIP-10 ............................ 500-P129bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat IP-10 .............................................. 500-P290 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat IP-10 .......................... 500-P290bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human I-TAC ....................................... 500-P132 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hI-TAC ............................. 500-P132bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine JE (MCP-1).............................. 500-P113 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mJE (MCP-1) .................. 500-P113bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine KC ........................................... 500-P115 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mKC ................................ 500-P115bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human KGF ......................................... 500-P19 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hKGF ............................... 500-P19bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human LD78β ..................................... 500-P187G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hLD78β ........................... 500-P187Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human LEC/NCC-4 ............................. 500-P125G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hLEC/NCC-4 ................... 500-P125Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Leptin ...................................... 500-P86 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hLeptin ............................ 500-P86bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-murine Leptin ...................................... 500-P68 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-mLeptin ........................... 500-P68bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat Leptin ............................................. 500-P185G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-rat Leptin ........................ 500-P185Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human LIGHT ..................................... 500-P179 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hLIGHT ........................... 500-P179bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine LIX .......................................... 500-P146 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mLIX ............................... 500-P146bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Lymphotactin .......................... 500-P40 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hLymphotactin ................ 500-P40bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Maspin .................................... 500-P270 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hMaspin .......................... 500-P270bt .............. 25 μg ..............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MCP-1/MCAF ......................... 500-P34 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hMCP-1/MCAF ............... 500-P34bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat MCP-1 ............................................ 500-P76 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat MCP-1 .......................... 500-P76bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MCP-2 ..................................... 500-P35 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hMCP-2 ........................... 500-P35bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine MCP-2 ..................................... 500-P127 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mMCP-2 .......................... 500-P127bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MCP-3 ..................................... 500-P37 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hMCP-3 ........................... 500-P37bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MCP-3 ..................................... 500-P37G ................ 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hMCP-3 ........................... 500-P37Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine MCP-3 ..................................... 500-P116G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mMCP-3 .......................... 500-P116Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human MCP-4 ..................................... 500-P04 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hMCP-4 ........................... 500-P04bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MCP-4 ..................................... 500-P04G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hMCP-4 ........................... 500-P04Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine MCP-5 ..................................... 500-P61 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mMCP-5 .......................... 500-P61bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-human M-CSF ..................................... 500-P44 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hM-CSF ........................... 500-P44bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine M-CSF ..................................... 500-P62G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mM-CSF .......................... 500-P62Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human MDC........................................ 500-P107 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hMDC ............................. 500-P107bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine MDC ....................................... 500-P176 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mMDC ............................ 500-P176bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MEC ........................................ 500-P297 ................. 50 ug ............. 100 ug ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hMEC .............................. 500-P297bt .............. 25 ug ............... 50 ug ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIA ......................................... 500-P243 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hMIA ............................... 500-P243bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIA-2 ...................................... 500-P255 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hMIA-2 ........................... 500-P255bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Midkine ................................... 500-P171 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hMidkine ......................... 500-P171bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIG ......................................... 500-P50 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hMIG .............................. 500-P50bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-1α .................................. 500-P38 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hMIP-1α ........................ 500-P38bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-1α .................................. 500-P38G ................ 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hMIP-1α ........................ 500-P38Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine MIP-1α ................................. 500-P121 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mMIP-1α ...................... 500-P121bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat MIP-1α ......................................... 500-P77 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat MIP-1α ..................... 500-P77bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-1β .................................. 500-P38B ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hMIP-1β ....................... 500-P38Bbt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine MIP-1β .................................. 500-P213 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mMIP-1β ...................... 500-P213bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine MIP-1γ .................................. 500-P117 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mMIP-1γ ....................... 500-P117bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N





Anti-murine MIP-2 ...................................... 500-P130 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mMIP-2 ........................... 500-P130bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-3 ...................................... 500-P124 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hMIP-3 ............................ 500-P124bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-3α ................................. 500-P95A................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hMIP-3α ....................... 500-P95Abt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-3β .................................. 500-P95B ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hMIP-3β ....................... 500-P95Bbt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-4 ...................................... 500-P108 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hMIP-4 ............................ 500-P108bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human MIP-5 ...................................... 500-P123G .............. 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hMIP-5 ............................ 500-P123Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Nanog ..................................... 500-P236 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hNanog ........................... 500-P236bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human NAP-2 ..................................... 500-P03 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hNAP-2 ........................... 500-P03bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human NAP-2 ..................................... 500-P03G ................ 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hNAP-2 ........................... 500-P03Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Neuroserpin ............................ 500-P271 ................. 50 μg ............. 100 μg ..............$2,100.................... rabbit ......................E, WB, NABiotinylated anti-hNeuroserpin .................. 500-P271bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human Neurturin ................................ 500-P102 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hNeurturin ...................... 500-P102bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human β-NGF ..................................... 500-P85 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hβ-NGF ........................... 500-P85bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human NNT-1/BCSF-3 ........................ 500-P186 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hNNT-1/BCSF-3 ............. 500-P186bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human NOV ........................................ 500-P257 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hNOV ............................. 500-P257bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human NP-1 ....................................... 500-P126G ............. 50 μg ............ 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hNP-1 ............................ 500-P126Gbt .......... 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human NT-3 ........................................ 500-P82G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hNT-3 .............................. 500-P82Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NA

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N




Anti-human NT-4 ........................................ 500-P83G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hNT-4 .............................. 500-P83Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Oct4-TAT ................................. 500-P289 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hOct4-TAT ....................... 500-P289bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Oncostatin M .......................... 500-P30 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hOncostatin M ................ 500-P30bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human OPG ....................................... 500-P149 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hOPG ............................. 500-P149bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human PAI-1 ....................................... 500-P260 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hPAI-1 ............................ 500-P260bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human PDGF-AA ................................ 500-P46 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hPDGF-AA ...................... 500-P46bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human PDGF-BB ................................ 500-P47 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hPDGF-BB ...................... 500-P47bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Persephin ................................ 500-P138 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hPersephin ...................... 500-P138bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human PF-4 ......................................... 500-P05 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hPF-4 .............................. 500-P05bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human PlGF-1 ..................................... 500-P226 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hPlGF-1........................... 500-P226bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-Human PTHrP .................................. 500-P276 ................ 50 μg ............. 100 μg ............. $2,100.................... rabbit ......................E, WB, NABiotinylated Anti-hPTHrP ........................... 500-P276bt ............ 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human sRANKL .................................. 500-P133 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hsRANKL ........................ 500-P133bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sRANKL .................................. 500-P133G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hsRANKL ........................ 500-P133Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine sRANKL .................................. 500-P63 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-msRANKL ....................... 500-P63bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sRANK Receptor ................... 500-P144 ................ 50 μg ............ 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hsRANK Receptor ............. 500-P144bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human RANTES .................................. 500-P36 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hRANTES ........................ 500-P36bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA


Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N



Anti-murine RANTES .................................. 500-P118 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mRANTES ....................... 500-P118bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat RANTES ......................................... 500-P78 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-rat RANTES .................... 500-P78bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine RELMα.................................... 500-P214 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-mRELMα ........................ 500-P214bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human RELMβ .................................... 500-P217 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hRELMβ .......................... 500-P217bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine RELMβ .................................... 500-P215 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-mRELMβ ......................... 500-P215bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Resistin .................................... 500-P183G .............. 50 μg ............. 100 μg ............. $2,100........................goat ..................... E, WB, NA, IBiotinylated Anti-hResistin ......................... 500-P183Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine Resistin .................................... 500-P182G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-mResistin ........................ 500-P182Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human SCF .......................................... 500-P48 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hSCF ............................... 500-P48bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human SCF .......................................... 500-P48G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hSCF ............................... 500-P48Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine SCF .......................................... 500-P71 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mSCF .............................. 500-P71bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat SCF................................................. 500-P202 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat SCF ............................ 500-P202bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human SCGF-β .................................. 500-P99G ................ 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hSCGF-β ........................ 500-P99Gbt ............ 25 μg ...............50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human SDF-1α .................................. 500-P87A................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hSDF-1α ........................ 500-P87Abt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine SDF-1α .................................. 500-P164G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mSDF-1α ....................... 500-P164Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human SDF-1β ................................ 500-P87BG .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hSDF-1β ...................... 500-P87BGbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine SF20 ........................................ 500-P259 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-mSF20 ............................ 500-P259bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NA



Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N



Anti-human TACI ........................................ 500-P166G .............. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hTACI .............................. 500-P166Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human TARC ....................................... 500-P97 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hTARC ............................. 500-P97bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TECK ....................................... 500-P134 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hTECK ............................ 500-P134bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TFF-2 ....................................... 500-P312 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hTFF-2 ............................ 500-P312bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TGF-α ................................... 500-P16 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hTGF-α ......................... 500-P16bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TIMP-1 .................................... 500-P280 ................. 50 μg ............. 100 μg ............. $2,100.................... rabbit ..................... E, WB, N, IBiotinylated Anti-hTIMP-1.......................... 500-P280bt .............. 25 μg ................50 μg ............. $4,000.................... rabbit ......................E, WB, NAAnti-human TL-1A .................................... 500-P240 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hTL-1A .......................... 500-P240bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TNF-α ................................... 500-P31A................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hTNF-α ......................... 500-P31Abt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TNF-α ................................... 500-P31AG .............. 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hTNF-α ......................... 500-P31AGbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine TNF-α .................................. 500-P64 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-mTNF-α ....................... 500-P64bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine TNF-α .................................... 500-P64G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-mTNF-α ......................... 500-P64Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-rat TNF-α ........................................... 500-P72 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-rat TNF-α ....................... 500-P72bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TNF-β ..................................... 500-P31B ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hTNF-β .......................... 500-P31Bbt ............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sTNF-Receptor Type I .................... 500-P143 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated Anti-hsTNF-Receptor TypeI ............ 500-P143bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human sTNF-Receptor Type II ................... 500-P168 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hsTNF-Receptor Type II ......... 500-P168bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TPO ......................................... 500-P49G ................ 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hTPO ............................... 500-P49Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NA


Polyclonal/Biotinylated Antibodies Antigen Affinity-PurifiedK

ey: E = ELISA, W

B = W

estern blot, N = N

eutralization, NA

= Neutralization data not available, I = IH

C


Anti-human sTRAIL/Apo2L ........................ 500-P135 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hsTRAIL/Apo2L .............. 500-P135bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-murine TRAIL ...................................... 500-P303 ................. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-mTRAIL ........................... 500-P303bt .............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human sTRAIL Receptor-2 .................. 500-P299 ................. 50 μg ............. 100 μg ............. $2,100........................goat .......................E, WB, NABiotinylated Anti-hsTRAIL Receptor-2 ........ 500-P299bt .............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human TSLP ........................................ 500-P258 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hTSLP ............................ 500-P258bt ............ 25 μg ...............50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human TWEAK ................................... 500-P137G .............. 50 μg ............. 100 μg ............. $2,100........................goat ........................ E, WB, NBiotinylated Anti-hTWEAK ........................ 500-P137Gbt ........... 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-human Vaspin ..................................... 500-P256 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hVaspin ........................... 500-P256bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human VCAM-1 .................................. 500-P300 .................. 50ug .............. 100ug ............. $2,100.......................rabbit ......................E, WB, NABiotinylated Anti-hVCAM-1 ........................ 500-P300bt ............... 25ug ................ 50ug ............. $4,000.......................rabbit ......................E, WB, NAAnti-human VEGF ...................................... 500-P10 ................... 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-hVEGF ............................ 500-P10bt ................ 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human VEGF ...................................... 500-P10G ................ 50 μg ............. 100 μg ............. $2,100........................goat ...................... E, WB, N, IBiotinylated Anti-hVEGF ............................ 500-P10Gbt ............. 25 μg ................50 μg ............. $4,000........................goat .......................E, WB, NAAnti-murine VEGF ...................................... 500-P131 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ..................... E, WB, N, IBiotinylated Anti-mVEGF ........................... 500-P131bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-rat VEGF ............................................. 500-P275 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit ....................... E, WB, NBiotinylated anti-rat VEGF ......................... 500-P275bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human Visfatin .................................... 500-P222 .................. 50ug .............. 100ug ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hVisfatin.......................... 500-P222bt ............... 25ug ................ 50ug ............. $4,000.......................rabbit ......................E, WB, NAAnti-human WNT-1 .................................... 500-P250 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hWNT-1 .......................... 500-P250bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NAAnti-human WNT-3A .................................. 500-P251 ................. 50 μg ............. 100 μg ............. $2,100.......................rabbit .................... E, WB, NA, IBiotinylated Anti-hWNT-3A ....................... 500-P251bt .............. 25 μg ................50 μg ............. $4,000.......................rabbit ......................E, WB, NA

Control AntibodiesControl Goat Immunoglobulin .................. 500-G00 .................. 1 mg ............$65.00Control Rabbit Immunoglobulin ................ 500-P00 ................. 500 μg .............$50.00Control Mouse Immunoglobulin ................ 500-M00 ................ 250 μg .............$65.00



Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N



Monoclonal Antibodies

Anti-human Adiponectin .......................... 500-M126 ...................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-human BAFF .................................... 500-M64 ......................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-human BMP-4 ................................. 500-M121 ...................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human BMP-7 .................................. 500-M123 ...................500 μg .......................... $150 ...........................mouse ...................... E, NA

Anti-human Eotaxin ............................... 500-M25 ......................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-human Eotaxin-2 ............................. 500-M31 ......................500 μg .......................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human Eotaxin-3 ............................. 500-M32 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-murine EGF ...................................... 500-M136 .....................500 μg ........................... $150 .............................. rat ....................... E, WB, N

Anti-human FGF-basic ........................... 500-M38 ......................500 μg .......................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human FGF-5 .................................. 500-M40 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human IFN-γ .................................... 500-M90 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human IL-2 ...................................... 500-M02 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-murine IL-2 ...................................... 500-M127 .....................500 μg ........................... $150 .............................. rat ....................... E, WB, N

Anti-rat IL-2 ............................................. 500-M129 .....................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human sIL-2R .................................. 500-M02R ....................500 μg ........................... $150 ...........................mouse ..................E, WB, N, I

Anti-human IL-4 ...................................... 500-M04 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human IL-6 ...................................... 500-M06 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human IL-7 ...................................... 500-M07 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human IL-8 ...................................... 500-M08 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human IL-10 .................................... 500-M86 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-murine IL-10 .................................... 500-M128 .....................500 μg ........................... $150 .............................. rat ....................... E, WB, N

Anti-rat IL-10 ........................................... 500-M130 .....................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human IL-11 .................................... 500-M11 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human IL-12 .................................... 500-M12 ......................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-murine IL-12 .................................... 500-M59 ......................500 μg .......................... $150 .............................. rat ....................... E, WB, N

Anti-human IL-15 .................................... 500-M15 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human IP-10 .................................... 500-M60 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-rat IP-10 ........................................... 500-M130 .....................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human LEC ...................................... 500-M43 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

PRoduCt CatalogdESCRIPtIon numBER SIzE PRICE SouRCE aPPlICatIonS

Continued on next page…

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N



PRoduCt CatalogdESCRIPtIon numBER SIzE PRICE SouRCE aPPlICatIonS

Monoclonal Antibodies

Anti-human Leptin ................................... 500-M27 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human MCP-1 .................................. 500-M71 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human MCP-2 ................................. 500-M69 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human MCP-3 .................................. 500-M73 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human MCP-4 .................................. 500-M70 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human MDC..................................... 500-M41 ......................500 μg .......................... $150 ...........................mouse ...................... E, NA

Anti-human MIG ..................................... 500-M42 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human MIP-1α ............................... 500-M74 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human MIP-3α ................................. 500-M28 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human MIP-3β ................................ 500-M29 ......................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-human NAP-2 .................................. 500-M33 .......................500 μg ........................... $150 ...........................mouse .................. E, WB, NA

Anti-human β-NGF .................................. 500-M85 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Anti-human NT-4 ..................................... 500-M24 .......................500 μg ........................... $150 ...........................mouse ...................... E, NA

Anti-human sRANKL ............................... 500-M46 .......................500 μg ........................... $150 ...........................mouse ...................... E, NA

Anti-human RANTES ............................... 500-M75 .......................500 μg ........................... $150 ...........................mouse ..................E, WB, N, I

Anti-human Resistin ................................. 500-M91 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human SCF ...................................... 500-M44 ......................500 μg .......................... $150 ...........................mouse .................. E, WB, NA

Anti-human TECK ................................... 500-M48 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human TGF-β1 .................................. 500-M66 .......................500 μg ........................... $150 ...........................mouse ................ E, WB, NA, I

Anti-human TNF-α .................................. 500-M26 .......................500 μg ........................... $150 ...........................mouse ..................E, WB, N, I

Anti-human sTRAIL/Apo2L ..................... 500-M49 ......................500 μg .......................... $150 ...........................mouse ................... E, WB, N

Anti-human VEGF ................................... 500-M88 .......................500 μg ........................... $150 ...........................mouse ................... E, WB, N

Control Mouse Immunoglobulin ............. 500-M00 ....................... 250μg .......................... $65.00

Key: E = ELISA

, WB

= Western blot, N

= Neutralization, N

A = N



ELISA Development Kits

Human 4-1BBR .............................. 900-K208 ....................... $320.00Human BD-1 .................................. 900-K202 ........................ $320.00Human BD-2 ................................ 900-K172 ....................... $320.00Human BD-3 ................................ 900-K210 ....................... $320.00Human BD-4 ................................ 900-K435 ....................... $320.00Human Betacellulin ...................... 900-K372 ....................... $320.00Human BMP-2 ............................. 900-K255 ....................... $320.00Human sCD40L ............................ 900-K145 ....................... $320.00Human CNTF ............................... 900-K158 ....................... $320.00Rat CNTF ...................................... 900-K65 ......................... $320.00Human CTGF ............................... 900-K317 ....................... $320.00Human CXCL16 ........................... 900-K230 ....................... $320.00Human EGF ................................. 900-K05 ......................... $320.00Murine EGF .................................. 900-K179 ....................... $320.00Rat EGF ........................................ 900-K390 ....................... $320.00Human EG-VEGF ......................... 900-K244 ....................... $320.00Human Eotaxin ............................ 900-K69 ......................... $320.00Murine Eotaxin ............................ 900-K68 ......................... $320.00Human Eotaxin-3 ......................... 900-K167 ....................... $320.00Human FGF-basic ........................ 900-K08 ......................... $320.00Human Follistatin ......................... 900-K299 ....................... $320.00Human G-CSF .............................. 900-K77 ......................... $320.00Murine G-CSF............................... 900-K103 ....................... $320.00Human GM-CSF ........................... 900-K30 ......................... $320.00Murine GM-CSF ........................... 900-K55 ......................... $320.00Human GRO/MGSA ..................... 900-K38 ......................... $320.00Rat GRO/KC ................................. 900-K57 ......................... $320.00Human GROβ................................. 900-K120 ....................... $320.00Human Heregulinβ-1 .................... 900-K316 ....................... $320.00Human ICAM-1 ............................ 900-K464 ....................... 4320.00Human IFN-γ ............................... 900-K27 ......................... $320.00

Murine IFN-γ ................................ 900-K98 ......................... $320.00Rat IFN-γ ...................................... 900-K109 ....................... $320.00Human IGF-BP1 ........................... 900-K315 ....................... $320.00Murine IGF-I ................................ 900-K170 ....................... $320.00Human IL-1α ................................ 900-K11 ......................... $320.00Murine IL-1α ................................ 900-K82 ......................... $320.00Rat IL-1α ...................................... 900-K204 ....................... $320.00Murine IL-1β ................................ 900-K47 ......................... $320.00Rat IL-1β ...................................... 900-K91 ......................... $320.00Human IL-1RA ............................. 900-K474 ....................... $320.00Human IL-2 .................................. 900-K12 ......................... $320.00Murine IL-2 .................................. 900-K108 ....................... $320.00Rat IL-2 ........................................ 900-K205 ....................... 4320.00Human IL-3 .................................. 900-K13 ......................... $320.00Murine IL-3 .................................. 900-K48 ......................... $320.00Human IL-4 .................................. 900-K14 ......................... $320.00Murine IL-4 .................................. 900-K49 ......................... $320.00Human IL-5 .................................. 900-K15 ......................... $320.00Human IL-6 .................................. 900-K16 ......................... $320.00Murine IL-6 .................................. 900-K50 ......................... $320.00Rat IL-6 ........................................ 900-K86 ......................... $320.00Human IL-7 .................................. 900-K17 ......................... $320.00Human IL-8 .................................. 900-K18 ......................... $320.00Human IL-9 .................................. 900-K20 ......................... $320.00Human IL-10 ................................ 900-K21 ......................... $320.00Murine IL-10 ................................ 900-K53 ......................... $320.00Human IL-11 ................................ 900-K22 ......................... $320.00Human IL-12 ................................ 900-K96 ......................... $320.00Murine IL-12 ................................ 900-K97 ......................... $320.00Human IL-13 ................................ 900-K23 ......................... $320.00

Each ELISA Kit contains the following components: Capture antibody, Detection antibody (biotinylated), Avidin Peroxidase Conjugate and a carefully calibrated Standard sufficient to assay the target cytokine in at least 1,000 ELISA plate wells.

Catalog PRICEPRoduCt dESCRIPtIon numBER PER KIt


Continued on next page…


Additional kits are under development.For the latest listing of products please visit our website at

www.peprotech.com

Murine IL-13 ................................ 900-K207 ....................... $320.00Human IL-17/IL-17A .................... 900-K84 ......................... $320.00Murine IL-17 ................................ 900-K392 ....................... $320.00Human IL-17E.............................. 900-K234 ....................... $320.00Human IL-17F .............................. 900-K277 ....................... $320.00Human IL-20 ................................ 900-K224 ....................... $320.00Human IL-21 ................................ 900-K226 ....................... $320.00Murine IL-21 ................................ 900-K368 ....................... $320.00Human IL-22 ................................ 900-K246 ....................... $320.00Human IL-31 ................................ 900-K347 ....................... $320.00Human IL-33 ................................. 900-K398 ........................ $320.00Human IP-10 ................................ 900-K39 ......................... $320.00Murine IP-10 .................................. 900-K153 ..................... $320.00Rat IP-10 ........................................ 900-K449 ........................ $320.00Human I-TAC ................................ 900-K151 ....................... $320.00Murine JE/MCP-1 ......................... 900-K126 ....................... $320.00Murine KC .................................... 900-K127 ....................... $320.00Human Leptin ............................... 900-K90 ......................... $320.00Murine Leptin ............................... 900-K76 ......................... $320.00Human MCP-1 ............................. 900-K31 ......................... $320.00Rat MCP-1 .................................... 900-K59 ......................... $320.00Murine MCP-3 .............................. 900-K123 ....................... $320.00Murine M-CSF .............................. 900-K245 ....................... $320.00Murine MDC ................................ 900-K197 ....................... $320.00Human MIA-2 .............................. 900-K357 ....................... $320.00Human Midkine ........................... 900-K190 ....................... $320.00Human MIG ................................. 900-K87 ......................... $320.00Human MIP-1α ............................ 900-K35 ......................... $320.00Murine MIP-1α ............................. 900-K125 ....................... $320.00Rat MIP-1α ................................... 900-K75 ......................... $320.00Murine MIP-1β .............................. 900-K278 ........................ $320.00Murine MIP-2 ............................... 900-K152 ....................... $320.00Human NAP-2 .............................. 900-K40 ......................... $320.00Human Neuroserpin ..................... 900-K412 ....................... $320.00

Human NOV ................................. 900-K338 ....................... $320.00Human β-NGF .............................. 900-K60 ......................... $320.00Human PAI-1 ................................ 900-K383 ....................... $320.00Human PDGF-BB ......................... 900-K04 ......................... $320.00Human PlGF-1 ............................. 900-K307 ....................... $320.00Human sRANKL ........................... 900-K142 ....................... $320.00Murine sRANKL ........................... 900-K233 ....................... $320.00Human RANTES .......................... 900-K33 ......................... $320.00Murine RANTES ........................... 900-K124 ....................... $320.00Rat RANTES ................................. 900-K72 ......................... $320.00Human Resistin ............................ 900-K235 ....................... $320.00Human SCF .................................. 900-K34 ......................... $320.00Murine SCF .................................. 900-K78 ......................... $320.00Rat SCF......................................... 900-K258 ....................... $320.00Human TACI ................................. 900-K216 ....................... $320.00Human TIMP-1 ............................ 900-K438 ....................... $320.00Human TL-1A ............................... 900-K290 ....................... $320.00Human TNF-α .............................. 900-K25 ......................... $320.00Murine TNF-α .............................. 900-K54 ......................... $320.00Rat TNF-α .................................... 900-K73 ......................... $320.00Human TPO.................................. 900-K44 ......................... $320.00Human sTRAIL/Apo2L ................. 900-K141 ....................... $320.00Human TSLP ................................ 900-K334 ....................... $320.00Human TWEAK ........................... 900-K149 ....................... $320.00Human VEGF ............................... 900-K10 ......................... $320.00Murine VEGF ............................... 900-K99 ......................... $320.00Rat VEGF ..................................... 900-K436 ....................... $320.00

ELISA Development Kits




Mini ELISA Development Kits

Human BD-1 ..................................900-M202 ........................ $100.00Human BD-2 ..................................900-M172 ........................ $100.00Human BD-4 ..................................900-M435 ........................ $100.00Human BMP-2 ...............................900-M255 ........................ $100.00Human sCD40 Ligand ...................900-M145 ........................ $100.00Human CTGF .................................900-M317 ........................ $100.00Human CXCL16 .............................900-M230 ........................ $100.00Human EGF ....................................900-M05 ......................... $100.00Rat EGF ..........................................900-M390 ........................ $100.00Murine Eotaxin-2 ............................900-M31 ......................... $100.00Human FGF-basic ...........................900-M08 ......................... $100.00Human GM-CSF .............................900-M30 ......................... $100.00Human GROβ ................................900-M120 ........................ $100.00Human Heregulinβ-1 .....................900-M316 ........................ $100.00Human ICAM-1 .............................900-M464 ........................ $100.00Human IFN-γ ..................................900-M27 ......................... $100.00Murine IFN-γ...................................900-M98 ......................... $100.00Human IL-1RA ...............................900-M474 ........................ $100.00Murine IL-1β ...................................900-M47 ......................... $100.00Human IL-2 ....................................900-M12 ......................... $100.00Rat IL-2 ..........................................900-M205 ........................ $100.00Murine IL-3 .....................................900-M48 ......................... $100.00Human IL-4 ....................................900-M14 ......................... $100.00Human IL-5 ....................................900-M15 ......................... $100.00Murine IL-5 ....................................900-M406 ........................ $100.00Human IL-6 ....................................900-M16 ......................... $100.00Murine IL-6 .....................................900-M50 ....................... $100.00Rat IL-6 ...........................................900-M86 ......................... $100.00Human IL-8 ....................................900-M18 ......................... $100.00Murine IL-10 ...................................900-M53 ......................... $100.00Human IL-11 ..................................900-M22 ......................... $100.00Human IL-12 ..................................900-M96 ......................... $100.00Murine IL-12 ...................................900-M97 ......................... $100.00

Murine IL-13 ..................................900-M207 ........................ $100.00Murine IL-15 ..................................900-M188 ........................ $100.00Human IL-17A ................................900-M84 ......................... $100.00Murine IL-17 ..................................900-M392 ........................ $100.00Human IL-17E ...............................900-M234 ........................ $100.00Murine IL-21 ..................................900-M368 ........................ $100.00Human IL-31 .................................900-M347 ........................ $100.00Human IL-33 .................................900-M398 ........................ $100.00Murine IP-10 ..................................900-M153 ........................ $100.00Murine JE/MCP-1 ...........................900-M126 ........................ $100.00Murine KC ......................................900-M127 ........................ $100.00Human MCP-1 ................................900-M31 ......................... $100.00Human MIG ....................................900-M87 ......................... $100.00Human MIP-1α ...............................900-M35 ......................... $100.00Murine MIP-1α ..............................900-M125 ........................ $100.00Murine MIP-1β ..............................900-M278 ........................ $100.00Murine MIP-2 ................................900-M152 ........................ $100.00Human Neuroserpin ......................900-M412 ........................ $100.00Human RANTES .............................900-M33 ......................... $100.00Murine RANTES ............................900-M124 ........................ $100.00Rat RANTES ...................................900-M72 ......................... $100.00Rat SCF ..........................................900-M258 ........................ $100.00Human TIMP-1 ..............................900-M438 ........................ $100.00Human TNF-α ................................900-M25 ......................... $100.00Murine TNF-α .................................900-M54 ......................... $100.00Human VEGF .................................900-M10 ......................... $100.00Rat VEGF .......................................900-M436 ........................ $100.00



Each Mini ELISA kit contains the following components: Capture antibody, Detection antibody (biotinylated), a calibrated Standard and Avidin-HRP Conjugate, sufficient to assay the target protein in at least 200 ELISA plate wells.

NEW

Additional kits are under development.For the latest listing of products please visit our website at

www.peprotech.com


Antigen Affinity Purification of Antibodies

Appendix I

(A.) Preparation of separation resin. (B.) Specific binding of Antibody to the resin. (C.) pH elution of the antibody from the affinity column.

Figure I-1 - Antigen Affinity Purification of AntibodiesCytokines are signaling proteins necessary for cell-to-cell communication throughout the body. In accordance with their role in preserving vital biological functions, the amino acid sequence of homologous cytokines from different mammalian species is general-ly highly conserved, rendering them poor immunogens for antibody production. Consequently, the sera obtained from host animals after immunization with cytokines (Figure I-4) typically contain minute amounts of cytokine-specific antibodies. Isolation of these antibod-ies by standard purification procedures (e.g. ion-exchange chroma-tography) is tedious and ineffective.

On the other hand, purification protocols which exploit the bind-ing affinity of antibodies to certain bio-molecules have yielded much better results. For example, Protein A (Staphylococcus-derived) and Protein G (Streptococcus-derived) are bacterial proteins that possess high binding affinity toward the Fc (Fragment crystallizable) region of immunoglobulins (IgGs), and when attached to a solid-support matrix, enable efficient separation of IgGs from other serum constit-uents. Protein A/G affinity purification of serum-IgGs typically en-riches the desired antibody by more than 100-fold. Yet, in the case of anti-cytokine antibodies, the total IgG fraction often contains less than 0.2% of the desired antibody. The large quantity of unrelated IgGs found in these preparations not only interferes with quantification of the relative amount of the cytokine-specific antibody, but also con-siderably increases the background noise when the antibody is used in analytical procedures such as ELISA, neutralization, immunohis-tochemistry, and Western blotting. A superior method for isolating specific polyclonal antibodies from antiserum is by antigen affinity chromatography, which exploits the specificity of antibody-antigen interactions (Figure I-1). Here, the separation medium consists of a


Appendix I continued

Figure I-2 Schematic illustration of column chromatography setup for antigen-affinity purification of antibodies.

solid-support resin to which the antigen is attached through stable covalent bonds. The immobilized antigen is then used in a column chromatography setup to selectively capture antigen-specific antibod-ies, while other serum proteins and unrelated immunoglobulins are washed away (Figure I-2). The antigen-bound antibodies can be

eluted from the column by an acidic solution (Figure I-3), which is promptly brought back to physiological pH to prevent acid-catalyzed antibody denaturation. This method typically yields >95% pure spe-cific antibodies (Figure 1-5).

NOTE: Elution is titrated with 1M NaOH to pH 7.2.



Figure I-5 - SDS-PAGE analysis of Rabbit anti-human IL-8 polyclonal antibodies (RbαhIL-8)

Lane 1: Molecular weight markers Lane 2: 2µg of RbαhIL-8 (unreduced)Lane 3: 2µg of RbαhIL-8 (reduced)

1 2 3

31.0

6.0

36.5

55.4

66.3

97.4

14.4

21.5

116.3

200.0

Approx.M.W. (kDa)

Day Procedure Antiserum Collected 1 Prebleed 3mL 1 Initial Immunization (250ug) - 21 Immunization (125ug) - 31 Titer Test Bleed 3mL 42 Immunization (125ug) - 49 Production Bleed 10-25mL 56 Production Bleed 10-25mL 63 Immunization (125ug) - 70 Production Bleed 10-25mL 77 Production Bleed 10-25mL 84 Immunization (125ug) - 91 Production Bleed 10-25mL 98 Production Bleed 10-25mL

Figure I-4 - General Immunization Schedule for Polyclonal Antibody Production in Rabbits

Figure I-3 - A typical Antigen Affinity Purification Chromatogram

Antigen Affinity Purification of Antibodies Continued


Antibody FAQ 1. Does PeproTech test for endotoxin in their antibodies?

PeproTech does test their antibodies for endotoxin using the kinetic LAL method. Please contact Technical Support for more information.

2. Which isotype are PeproTech’s polyclonal antibodies?

The polyclonal antibodies that PeproTech manufactures are pre-dominantly IgG antibodies.

3. HowarePeproTech’santibodiespurified?

All polyclonal and biotinylated polyclonal antibodies are antigen-affinitypurified.Monoclonalantibodypurificationvariesbyproduct; please contact Technical Support for more information.

4. Can you tell me what epitope PeproTech’s antibody binds to?

PeproTech does not perform epitope mapping at this time. As a general guideline, a polyclonal mixture of antibodies will bind to multiple epitopes on the protein of interest while each monoclo-nalantibodywillbindtoaspecificepitope.

5. HavePeproTech’santibodiesbeentestedinneutralizationasays?

Neutralization testing is performed on a lot-to-lot basis for each antibody. The results of this testing can be found on the product’s corresponding data sheet.

6. Are PeproTech’s antibodies suitable for use in ELISA and Western Blot applications?

Most of PeproTech’s antibodies are suitable for use in ELISA and WesternBlotassays;pleaseseeourindividualELISAandWesternBlot F.A.Q. sections for more information regarding these applica-tions.

7. What information should be known about the stability of PeproTech’s antibody products?

PeproTech’s antibodies are lyophilized from PBS. As such, they are stable at room temperature for at least 1 month. For longer periods, we recommend storing the lyophilized products at -20°C.

For reconstituted solutions of the antibodies, we recommend short-term storage at 4°C. For long-term storage the antibody solu-tionshouldfirstbealiquoted(toavoidmorethanonefreeze/thawcycle) and stored frozen at -20°C.Frozenaliquotsofthisantibodysolution are stable for at least 1 year when kept at -20°C.

8. Do PeproTech’s antibody products contain any carrier proteins or otheradditives?

No, PeproTech does not formulate its polyclonal, biotinylated polyclonal,ormonoclonalantibodieswithadditivesorcarrierproteins.

9. Will PeproTech’s antibodies work in immunohistochemistry and immunocytochemistry applications?

Allantibodiesthathavebeentestedsofarhavebeenfoundtobesuitablefortheseapplications.However,wearestillgatheringdata for more of our antibodies. Please contact Technical Support for more information.

10. Will PeproTech antibodies recognize target proteins sold by other vendors?

PeproTech’santibodyproductshavehighbindingaffinitytowardsthenaturaland/orrecombinantversionsofthecorrespondingproteins.However,duetolackofauthenticitysometimesfoundinothervendors’proteins,wecannotguaranteethatourantibodieswill perform as well with these proteins.

11. Will PeproTech’s antibodies recognize target protein in complex biologicalfluidssuchasbloodorserum?

Yes.However,forsamplesthathaveahighcontentofinterferingagentstherecognitionwillbelessefficientandahigherback-ground, or reduced signal to noise ratio, may be seen.



Appendix II

ELISAEnzyme-linked immunosorbent assay (ELISA) is a laboratory

technique for quantifying the concentration of specific antigens or antibodies in cell lysates, sera, or other complex solutions. In its simplest format, called direct ELISA, a solid-phase binding support such as a polystyrene plate is used to capture the tested antigen, which is then detected with the aid of a specific enzyme-linked an-tibody (E-Ab). The detection protocol involves conversion of a col-orless substrate solution into an intense, colored product that can be measured directly in the reaction plate. The increase in optical density during the reaction is recorded at different time points for several sample dilutions, and the collected data is then used in con-junction with a standard curve to calculate the concentration of the antigen in the original sample (Figure II-1). Although the sensitivity and specificity of ELISA are comparable to those of radioimmuno-assay (RIA), ELISA has the advantage of not requiring radioactive isotopes and expensive counting devices. In recent years, ELISA has gained popularity in both research and clinical laboratories, and has become an important diagnostic test for many infectious dis-eases, including the human immunodeficiency virus (HIV).

A number of different enzymes are suitable for the preparation of E-Abs, including alkaline phosphatase, horseradish peroxidase, and carbonic anhydrase. Although E-Abs can be obtained by co-

valently binding enzymes to antibodies through a coupling agent such as glutaraldehyde, this method is not commonly used, because it often yields products with impaired immunological and/or enzy-matic activity. A superior method to prepare fully active E-Abs is by exploiting the high binding affinity of avidin for biotin, as depicted in Figure II-2. The biotinylated antibody used in this method con-tains on average 6-8 biotin moieties, which due to their small size do not interfere with the interaction between the antibody and the captured antigen. Once unbound antibody has been rinsed from the plate, the biotin moieties are used in-situ to form stable complexes with avidin-enzyme conjugates. A major limitation of the direct ELISA format is that certain antigens (e.g. low molecular weight cytokines) bind poorly to polystyrene plates, and therefore, cannot be measured accurately by the assay. To bypass this limitation, the plate can be pre-coated with a specific antibody that recognizes the tested antigen through epitopes that are not used by the E-Ab. Us-ing a matched pair of antigen-specific antibodies, one for capturing and one for detection, significantly improves both the sensitivity and specificity of the assay. This format, called Sandwich ELISA, enables detection of picogram quantities of antigens and is the pre-ferred method for measuring cytokines and hormones in biological samples (Figure II-3).


Appendix II continued

Figure II-1 - Sample Standard Curve - Human EGF ELISA



(A.) Biotinylation of an Antibody. (B.)Avidin-EnzymeConjugation.(C.) In Situformationofenzyme-linkedAntibodyvianoncovalentinteractionbetweenAvidinandBiotin.

Figure II-2 - Steps in the production of enzyme-linked antibody, through Avidin-Biotin interaction.

O



Figure II-3 - Schematic presentation of Sandwich ELISA protocol.

B BB

BBB BB

BB B B

BB

B

A. Prepare fresh solution of antigen-specificantibody (capture antibody) in PBS and promptly add 100µlaliquotstoeach well. Seal the plate and incubate at room temperature overnightorat37˚Cfor 2-4 hours. When incubation time is complete, wash away unbound antibody (i.e. aspirate and wash 4x).

Note:Unlessotherwisespecified,theELISAprotocolisconductedatambienttemperature.Amoredetailedprotocolmay be found on our website under the Protocols section.

B. Add fresh solution of blocking agent (i.e. BSA) and incubate for at least 1 hour. When incubationisover,wash away the blocking agent.

Note:Inefficientblocking of non-specificbinding may lead to high background noise. If thisisobserved,increase the incubation time or try other blocking agents.

C. Add freshly prepared solutions of standards, samples or controls and incubate for > 2 hours. Wash the wells 4x.

D. Add freshly prepared solution of biotinylated antigen-specificantibody(detection antibody), and incubate for 2 hours. Wash the wells 4x.

E. Add a solution of avidin-peroxidaseconjugateand incubate for 30 minutes. Wash the wells 4x.

F. Add substrate (ABTS) solution and readthedevelopingoptical density at 5 minuteintervalsusing an ELISA plate reader set at 405nm with a 650 nm wavelength correction.

Note: Blank optical density (O.D.) read-ings should not ex-ceed 0.2 O.D. units when using ABTS.


ELISA FAQ1.HowmanyplatescanIrunwithoneofPeproTech’sELISADevel-

opment Kits (EDKs)?

One EDK contains enough material to run ten 96 well plates, while one Mini EDK contains enough material to run two 96 well plates.

2.Whatisthestabilityoftheavidin-HRPincludedinPeproTech’sELISADevelopmentKit?

Theavidin-HRPincludedinthekitisstableforupto1monthat2-8˚C,andforupto2yearsat-20˚C.

3.HowcanIfindcrossreactivityinformationforakit?

PeproTechperformsin-housecrossreactivitytestingonitsELISADevelopmentKits.Theresultsthathavebeencollectedfromthistesting are located on the kit’s data sheet.

4. IsthereanystepoftheELISAprotocolthatcanbeleftovertheweekend?

The plate may be coated with the capture antibody on Friday, left at4˚Covertheweekend,andresumedonMonday.Please note: changing incubation times may cause results to vary between plates.

5. Are PeproTech’s EDKs suitable to use with all sample types?

AlthoughPeproTechhasnottestedallofitskitsineverymatrixavailable,theyshouldbesuitableforusein,butnotlimitedto:serum,plasma,cellculturesupernatant,urineandsaliva.

6. Is a stop solution necessary to stop the reaction?

Astopsolutionisnotneededwhenusingavidin-HRP+ABTS.Ingeneral,reliablestandardcurvesareobtainedwheneitherO.D.readings do not exceed 0.2 units for the zero standard concentra-tions, or 1.2 units for the highest standard concentration.

If a stop solution is desired, 1% sodium dodecyl sulfate (SDS) may be used to end the reaction. Stop solutions are not used in PeproTech’s laboratory, though literature has shown that SDS is the optimal stop solution for ABTS.


7. Can I use TMB with PeproTech’s EDKs?

PeproTech’s EDKs are optimized using ABTS and are therefore best usedinconjunctionwiththissubstrate.ThekitcanstillbeusedincombinationwithTMB,butonlyaftersomeadjustmentshavebeenmade:

•Theavidin-HRPprovidedinthekitcannotbeusedwithTMB;streptavidinmustbepurchasedseparately.

•Dilutionsofstreptavidinwillneedtobeoptimized. •Astopsolutionisgenerallyneededwhenusingstreptavidin+

TMB. Refer to manufacturer’s data sheet. •TheTMBreactiontime,priortotheadditionofstopsolution,

will need to be optimized. •Theplateistobereadat450nmwithacorrectionat650nm

when using recommended plates. 8. Why is D-mannitol added to the EDK components?

D-mannitol is added to the EDK components in order to aid in protein/antibodyvisualization.ItdoesnotalterELISAresults.

9.CanIusethecurveontheEDKdatasheetasmystandardcurve?

AseparatestandardcurvemustberunoneachELISAplate.Inotherwords,thecurvefromoneplatecannotbeusedforadiffer-entplate.ThecurvethatPeproTechprovidesontheEDKdatasheetisfordemonstrationpurposesonly,asachievedinPeproTech’slaboratory.

10.HowdoesPeproTechgeneratetheirstandardcurve?

WhenanELISAisruninPeproTech’slab,aMolecularDevices® plate reader and the SOFTmax® PRO software are used. This pro-gramusesthevaluesthatarereceivedandgeneratesa4-parametercurve.Hereistheequationthattheprogramuses:

4-PFit: x =concentration(pg/ml) y = O.D. (405nm – 650nm) A, B, C, & D correspond to the 4 parameters.*

*For more detailed information regarding the parameters, please contact the Technical Support Department.


ELISA Troubleshooting GuideWhen trouble shooting, prepare all fresh buffers. Sterile filter buffers containing BSA and allow all reagents to reach room temperature prior to use.

Problem Possible Source Suggestion

High Background/ Non-specificbindingofantibody •Modifyblockingbufferused(e.g.substitute Non-specific Color casein for BSA). Do not wash after blocking Development step; dump, blot & go directly to the next step.

Platenotwashedsufficiently •Besureallwellsarefilledwithbufferduring everywashstep.Besurewashingapparatusis working properly. Wash 3-5x between steps. Donotaddadditionalwashes.Afterfinalwash, blotplateforcefullyonpapertoweltoremove residual buffer. Be sure the correct amount of Tween-20 was added to the wash solution (0.01- 0.1% recommended).

ContaminatedBuffers •Prepareallbuffersfresh(within7daysofuse) andsterilefilter.

Incubationtemperaturetoohigh •Incubateatroomtemperature(25ºC)throughout whole procedure.

Incubationtimetoolong •Reduceincubationtime.

Contaminationofthesubstrate •Changepipettetipsbetweenreagentsanduse withtheconjugateorcontamination separatereservoirs.Ideally,washbuffer ofblankwellswithpositivecontrol shouldbeaspirated(pouring/dumpingmaylead to cross contamination).

Concentrationofdetectionantibody •Checkcalculationsortryfurtherdilutions. oravidin-HRPistoohigh

Substrateexposedtolightpriortouse •Keepsubstrateinthedarkuntilreadyto dispense into wells.

Wrongfilterusedwhentakingreadings •Wavelengthshouldbe405nmforABTS(450for TMB)witha650nmwavelengthcorrection when using recommended plates.

Readbeyondtimeneededfordevelopment •Readat5minuteintervalsinordertomonitor blank O.D. readings.


Continued…



Weak/No Color Areagentorastepoftheprocedure •Checkprotocolandfollowstepscarefully.Development omitted by mistake

Detergentconcentrationinwashbuffer •Removeordecreasedetergent(0.01-0.1% too high recommended).

Platewashedinadequately •Decreasenumberofwashesbetweensteps, especially if not using automated or semi- automatedwasher.1-2washesmaysuffice.

Enzymeinhibitorpresentinsamples •Sodiumazideinhibitsperoxidaseactivity.

Wrongincubationtimeortemperature •Checkandfollowprotocolrecommendations. Place plates in an incubator during incubation periodstoavoidtemperaturefluctuations(setto 25ºC).

Wrongsubstratevolumeaddedtowells •Checkpipette;usecalibratedpipettes.

Wrongenzyme-substratesystemused •Checkmanufacturerspecifications:avidin-HRP +ABTS/streptavidin+TMB.

Conjugateorsubstrateinactive •Testactivityinanothersystemandcheck expiration dates. Incorrect storage of components •Storeallcomponentsasrecommendedondatasheet.

Wrongfilterusedwhentakingreadings •Wavelengthshouldbe405nmforABTS(450for TMB)witha650nmwavelengthcorrection when using recommended plates.

Poor Standard Curve Incorrectpreparationofstandard •Reconstitutestandardassuggestedondata sheet. Dilute only with recommended diluent bufferunlessalternativebufferhasbeenoptimized.

Dilutionsmadetooearly •Preparereagentsimmediatelypriortouseand add to wells promptly.

CaptureAntibodydidnotbind •UseELISA-suitableplates(i.e.NuncMaxiSorp Prod. # 439454); not tissue culture plates.

Inefficientwashing •Besurewashapparatusisworkingproperly(i.e. distributingevenvolumesintoeachwell).Be sure wells are empty after aspiration, yet be suretofillwellsinatimelymanner.

Pipettingerror •Dispensequicklyandidenticallyintothesideof each well. Use calibrated pipettes.


ELISA Troubleshooting Guide Continued




•Ingeneral,reliablestandardcurvesareobtained Readbeyondtimeneededfordevelopment when O.D. readings do not exceed 0.2 for blanks or 1.2 for the highest standard concentration.

Incorrectstorageofcomponents •Storeallcomponentsasrecommendedondata sheet. Do not allow reconstituted reagents to stay at room temperature for excess time.

Poor Precision Insufficientmixingofreagents •Ensureadequatemixingofreagentsbeforepipetting.

Inefficientwashing •Besurewashapparatusisworkingproperly(i.e. distributingevenvolumesintoeachwell).Be sure wells are empty after aspiration, yet be suretofillwellsinatimelymanner.

Pipettingerror •Dispensequicklyandidenticallyintothesideof each well. Use calibrated pipettes.

Reusedmaterials •Changepipettetipsbetweensamples,reservoirs between reagents, and plate sealers between incubation periods.

Wellshavebeenscratchedbypipette •Usecautionwhendispensingandaspirating. tips or washing apparatus

Precipitateinsamples •Centrifugevialspriortouse.

Uncleanwellsordebris •Inspectwellsandremovedebrispriortouse. Wipebottomofplatetoremoveanydebrisor fingerprintspriortoreading.

Edge Effects Evaporation •Useplatesealersbetweeneachstep.

Uneventemperature •Placeplatesinanincubatorduringincubation periodstoavoidtemperaturefluctuations (setto25ºC).Donotstackplates.

Poor Standard Curve (continued)


Appendix III

Western Transfer (Western Blot)

Typical Western Transfer and Development Protocol

Materials Needed• SDS-PAGE apparatus and accessories• Submerged Western Transfer Cassette and accessories• A nitrocellulose membrane, approximately the size of

the gel, presoaked in Western Transfer Buffer* for five minutes. (Note: The membrane should be handled with gloves and clean forceps to avoid contamination with extraneous proteins.)

• *Western Transfer Buffer (1/2 x TOWBIN) Tris Base ..........1.45g (12mM final concentration) Glycine ............7.2g (96mM final concentration) Methanol ..........200mL (20% final volume) diH2O ...............add to 1.0L final volume• Antigen specific probing antibody (Primary Antibody)• Secondary antibody (Donkey-anti-probing antibody

species conjugated to Alkaline Phosphatase)• Commercially available alkaline phosphatase conjugate

substrate kit

Figure III-1 - Schematic presentation of Western Transfer Protocol

Western Transfer, also known as Western Blotting, is a rapid immunoblotting technique for identifying the presence of a particu-lar protein in a complex mixture of proteins such as cell lysates or sera. The technique exploits both the efficiency of SDS-PAGE to separate a mixture of proteins into distinct protein bands, and the ability of immunochemical reagents to interact specifically with a given protein.

A typical Western transfer protocol (e.g. for the detection of protein X) includes the following steps: (1) The proteins in the test-ed solution are separated into distinct bands by SDS-PAGE, (2) The

size-separated proteins are transferred from the polyacrylamide gel to a nitrocellulose membrane (or another suitable matrix). (3) The membrane containing the protein bands is serially incubated with: (a) a suitable blocking reagent to prevent non-specific protein bind-ing, (b) a wash solution to rinse any unbound blocking reagent, (c) a probing antibody (anti-protein-X antibody) that forms a specific immune complex with Protein-X, (d) additional wash solution to remove any unbound antibody, (e) an enzyme-linked antibody that binds specifically to the Fc region of the anti-Protein-X antibody, (f) additional wash solution to remove any unbound enzyme-linked antibody, and finally, (g) a substrate solution, which in the presence of the enzyme, yields an insoluble, colored product that precipitates at the site of the immune complex, thereby rendering the Protein X band visible (Figure III-1).


Appendix III continued

Transfer Protocol1. Load the protein sample onto a 4-20% Tris-Glycine polyacryl-

amide gel and run until desired resolution is achieved. (The electrophoresis can be followed by using pre-stained molecular weight markers).

2. Set up the Submerged Western Transfer Cassette as described below and in Figure III-2:

a. Submerge the open Transfer Cassette cathode plate onto the tray pre-filled with Western Transfer Buffer.

b. Place a sponge support pad onto the cassette and remove the air bubbles by gently rolling a Pasteur pipette over the pad.

c. Place a piece of blotting paper onto the sponge support pad.d. Remove the gel from the electrophoresis plates, cut approxi-

mately 3mm off the bottom of the gel so that the membrane can be laid flat against the gel, and place the gel over the blotting paper. Expel air bubbles as before.

e. Carefully place the pre-soaked nitrocellulose membrane onto the gel and expel air bubbles. Ensure that the mem-brane remains directly over the gel before proceeding.

f. Place a second piece of blotting paper onto the nitrocellu-lose membrane and remove air bubbles.

g. Place a sponge support pad onto the second piece of blotting paper and remove air bubbles.

h. Gently close the cassette by placing the anode plate over the exposed pad.

3. Carefully place the assembled cassette into the transfer tank containing Western Transfer Buffer up to the “pre-fill” level and adjust the buffer level, as needed, after the addition of the cas-sette.

4. Connect the assembled apparatus to an electrophoresis power supply and run for approximately 1.5 hours at a constant current of 400mA.

Development (Immunostaining) Protocol1. After the transfer is complete, incubate the membrane in block-

ing solution for 30 minutes with gentle agitation on an orbital shaker. Note: Various types of blocking solution may be used depending on numerous assay variables; e.g. 3% Nonfat Dry Milk in diH2O or a fish-derived blocking buffer diluted in diH2O.

2. Wash the membrane three times with TBST (TBS, pH 7.2 with 0.1% TWEEN-20) in a clean tray on an orbital shaker; each wash lasting 5-10 minutes with fresh buffer.

3. Dilute the probing (primary) antibody in TBST to a volume of 50ml (approximate final concentration of 0.20µg/ml) and incu-bate the membrane in the antibody solution for one to four hours at room temperature. (The optimum incubation time depends on the antibody/antigen binding affinity and must be pre-determined for each antibody.)

4. Wash the membrane three times as in step #2.5. Dilute the enzyme conjugated secondary antibody in TBST

according to the manufacturer’s specification. Incubate the membrane in a clean tray containing 50ml of diluted secondary antibody for one hour at room temperature on an orbital shaker.

6. Wash the membrane three times as in step #2.7. Color development requires the use of a commercially available

(e.g. Bio-Rad or Sigma) alkaline phosphatase conjugate substrate kit. Follow the manufacturer’s instructions.

8. After the bands become clearly visible, stop the color develop-ment by placing the membrane in a tray containing diH2O for at least ten minutes.

Figure III-2 - Submerged Western Transfer Cassette

Transfer


1. How long will the immunostaining take when PeproTech’s Western Transfer protocol is followed?

The Western Transfer process will take approximately 6 hours fromtransferofproteinstovisualizationofbands.

2. What type of molecular weight marker should be used?

PeproTechusesNovex® Sharp Protein Standard pre-stained molecular weight marker for use in all Western Transfers performed in-house, although this molecular weight marker doesnothavetobeutilized.However,itisnecessarythatapre-stained molecular weight marker is used when not utiliz-ing the ECL detection method.

3. IstheadditionofapositivecontrolnecessaryinaWesternTransfer?

Yes, in order to know exactly how the protein of interest will visualizeontheWesternTransferyoumustaddapositivecontrol to your gel. When using PeproTech’s antibodies in a Western Transfer, we recommend using the corresponding PeproTech recombinant protein, which was the immunizing antigen for the antibody of choice.

4. DoesthecolordevelopmentsystemrecommendedinPepro-Tech’sWesternTransferprotocolhavetobeused?

No.TherearemanydifferentcolordevelopmentsystemsthatcanbeusedforthevisualizationofyourWesternTransfer.However,thesystemthatischosenmustbecompatiblewiththeenzymeconjugatebeingused.PeproTechusesanalkalinephosphataselinkedsecondaryantibody.NBT/BCIPis suitable for use with this enzyme, and is therefore used for thevisualizationofourWesternTransfers.

Western Transfer FAQ5. Is agitation of the membrane essential during the incubation

periods?

Yes, it is essential that the membrane be agitated during the incubation periods. If the membrane is not agitated the antibodies, blocking buffer, and washing buffer may not af-fectthemembraneevenlyandcancreatesplotchy,orpatchy,background. It can also limit the detection of the proteins by the antibodies.

6. Why are some Western Transfer results stronger than others when using different lots of the same antibody?

Duetothenatureofpolyclonalantibodies,variabilitymaybe seen from lot to lot.

7. Isitnecessarytoincludethefilterpaperduringtheproteintransfer step?

Yes, it is necessary to include the blotting paper as a barrier in the protein transfer step. This blotting paper helps protect the gel and membrane from any possible damage resulting from direct contact with the sponges, yet does not interfere with the electric current.

8. Cananenzyme-conjugatedprimaryantibodybeusedinaWesternTransferratherthanusingaprimary/secondaryanti-body system?

Yes,anenzyme-conjugatedprimaryantibodycanbeusedinsteadofutilizingaprimary/secondaryantibodysysteminyourWesternTransfer.However,byusingalabeledsecond-aryantibodythatrecognizestheantigenspecificprimaryantibody,therewillbeanamplificationofthesignalseenin a Western Transfer when compared to using the enzyme-conjugatedprimaryantibodyalone.



Problem Possible source Suggestion

Weak/No Signal Insufficient amount of protein • Increaseamountofproteininsample. present •Confirmproteinispresentthroughanalternate method.

Gel ran too long •Decreaserunningtimeandmonitorthegelby watching dye front and ensuring the current is turned off when dye front is near bottom of gel.

Old or degraded protein samples •Testafreshsampleoftargetprotein.

Protein over-transferred/ •Useamembranewithasmallerporesize. did not transfer correctly •DecreasevoltageforlowerMWproteins(less than 10kDa). •Makesuremembraneispre-soakedaccording to manufacturer’s instructions. •Checkandoptimizetransfertime,as appropriatetransfertimewillvarywithMWof the target protein. •Makesuretransfer“sandwich”issetupcorrectly andthereissufficientcontactbetweengeland membrane. (Please see our Western Transfer protocolforcorrecttransfer“sandwich”setup.) •Ensurethecorrecttransferbufferisbeingused. •ThepresenceofSDSduringtransfercanreduce the binding of the protein when using a nitrocellulose membrane. •Add20%methanoltotransferbuffertoincrease binding. •Checkproteintransferbystainingthemembrane withareversiblestain,suchasPonceauS,to visualizemajorbands.

Protein has a pI greater than 9 •SubstitutebufferwithahigherpHbuffer(i.e. CAPS buffer, pH 10.5).

Protein masked by blocking buffer •Testusingadifferentblockingbuffer. •Optimizeconcentrationofproteininblocking buffer. We recommend 3% Non-Fat Dry Milk in diH2O.


Western Transfer Troubleshooting Guide

Continued…




Weak/No Signal Enzyme inhibitor present (i.e. azide) • Make sure buffers do not contain sodium azide, (continued) as this will interfere with the HRP signal.

Old or degraded primary antibody •Checkmanufacturer’srecommendationsfor storageand/orexpirationdate.Ifexpiredorpast storage time, purchase new antibody. • Ifantibodywassubjectedtorepeatedfreeze/thaw cycles purchase new antibody, as this can affect thestructureandprotein:antibodybinding. •PerformaDotBlottoensureantibodyisstillactive.

Insufficient amount of primary • Check manufacturer’s recommendation for and/or secondary antibody antibody concentration. •Testarangeofconcentrationstofindtheoptimal conditionforindividualassay.

Incorrect primary antibody •Ensureprimaryantibodyreactswiththetarget protein from the species being studied (i.e. Rabbit-anti-human VEGF Human VEGF).

Incorrect secondary antibody •Ensurethesecondaryantibodybeingusedis against the species of the primary antibody (i.e. Donkey-anti-rabbit IgG Rabbit-anti-human VEGF).

Insufficient incubation • Increaseincubationperiodofprimaryantibody. of primary antibody Antibody can be incubated for at least one hour atroomtemperatureorovernightat4°C.

Insufficient incubation • Increase incubation period of secondary antibody of secondary antibody (please see manufacturer’s recommendations for incubation time and temperature).

Membrane over-washed •Shortenwashtimesand/orreducenumberofwashes. •Useawashingbufferthatdoesnotcontaina detergent, or that has a lower concentration of detergent present.

Incompatible developing reagent •Makesurethedevelopingreagentbeingusedis compatiblewiththeenzymeconjugate. •Ensuresubstrateismadecorrectlybyfollowing the manufacturer’s instructions.

Excess salt present •Decreaseamountofsaltpresentinwashbuffer. •Decreaseamountofsaltpresentinantibody dilution buffer.

Western Transfer Troubleshooting Guide Continued




Weak/No Signal Degraded/Incorrect developing • Ensurethedevelopingreagentwasprepared continued reagent correctly.

•Checkthatthereagentisstillactive.Ifnot, purchase new reagent.

For ECL method, film is bad/expired •Ensurefilmisnotexpiredorexposed.Ifso, purchasenewfilm.

For ECL method, ECL developing •UsefreshECLdevelopingsolutions. solutions are old/expired

For ECL method, film exposure • Increaseexposuretimeofthefilm. time is too short

For ECL method, cling film •Usedifferentbrandofclingfilm. interfering with reaction

High Background Antibody concentration too high • Check manufacturer’s recommendations for primaryand/orsecondaryantibodyconcentrations. •Testarangeofconcentrationstofindoptimal conditionforindividualassay.

Over-incubation of antibodies •Checkmanufacturer’sprotocoltoensurethe correct incubation time and temperature is used. If this information is not supplied by the manufacturer, optimize incubation time through testing.

Non-specific binding of antibodies •Ensuretheprimaryantibodyisspecificonlyfor the protein that is being targeted. •Runacontrol,omittingtheprimaryantibody incubation.Ifbandsdevelop,chooseadifferent secondary antibody. •Chooseadifferenthostspeciesforsecondary antibody.

Continued…




High Background Cross reaction of antibodies with • Use a different blocking buffer formulation. continued proteins in blocking buffer •GoatantibodieswillreactwithBSAduetoa and/or sample slightrecognitionofthebovineproteinbythe goat antibody. This will lead to a purple sheen on your membrane. If this occurs, use a different species antibody or a different blocking buffer. • Ifsecondaryantibodyissuspected,dilute antibody in buffer containing 1-5% normal serum from the same species as target protein.

Ineffective blocking buffer • Use a different blocking buffer formulation. •Adddetergenttoblockingbuffer,suchas TWEEN-20.

Insufficient blocking •Useadifferentblockingbufferformulation. •Batchofblockingbuffermaybeinactive.Use fresh blocking buffer. • Ifblockingovernightat4°C,thismaydecrease blockingefficiencysincelowertemperaturesmay lessentheeffectiveness.Tryincubationforone hour at room temperature. •AddTWEEN-20toblockingbuffer. •Optimizeincubationtimeand/ortemperatureof blocking step.

Insufficient washing •Aftereachstep,washmembrane3timesfor10 minutes each. Increase the duration of the washingstepifthisisnotsufficient. •Tryadifferentdetergentinwashingbufferoradd TWEEN-20 if not already present. • Increasevolumeofwashingbufferused.

Over-incubation in color •Shortenincubationtimeincolordevelopment development substrate solution.Monitordevelopmentuntilbandsare present and stop reaction by washing membrane in diH2O.

Excess enzyme present •Reduceconcentrationofselectedenzymeconjugate.

Interference by endogenous enzyme •NonFatDryMilkcontainsendogenousbiotinand mayinterferewithavidin/biotindetectionsystem. Use different blocking buffer, such as 3% BSA.

Contaminated reagents • Filterbuffersbeforeusetoremovecontaminant. •Makefreshbuffersandre-runWesternTransfer.





High Background Membrane problems •Testnewmembranes. continued •Duringtheentireprocedurethemembranecan only be dried between transfer and immunostaining. Once immunostaining has begun, the membrane cannot be dried until completion of immunostaining procedure. Re-run Western Transfer if membrane was dried at any other point during immunostaining procedure.

For ECL method, ECL film •Shortenfilmexposuretime. overexposed • Ifsignalfromtargetproteinistoostrong,wait5- 10minutesandre-exposetofilm.

Splotchy background Uneven agitation during incubations •Ensurethemembraneisagitatedevenlyby placing on a rocker or shaker during incubation.

Insufficient amount of incubation •Ensureenoughsolutionispresentduringeach buffer incubation period to fully submerge the membraneandallowittofloatfreelyinthe solution.

Air bubbles present during transfer •Gentlyremoveallairbubblesfromtransfer “sandwich”beforetransferisstarted.Thiscanbe donebyusingarollerorcleanglassrod/Pasteur pipette.

Membrane problems •Ensuremembraneiswettedthoroughlyaccording to the manufacturer’s protocol. •Handlemembraneswithextremecare. Mishandledmembranescancausenon-specific binding. •Handlingmembraneswithbarehandscanleadto contamination of membrane. Use forceps or wear gloveswhenhandlingmembrane.

Aggregation of HRP conjugate • IfusingHRP,filterconjugatetoremoveHRP aggregates. •UseafreshsampleofHRPconjugate.

Contamination of reagents •Filterbuffersbeforeusetoremovecontaminant. •Makefreshbuffersandre-runWesternTransfer.

Continued…




Non-specific bands Antibody concentration too high •Checkmanufacturer’srecommendationsfor primaryand/orsecondaryantibodyconcentrations. •Testarangeofconcentrationstofindoptimal conditionforindividualassay.

Old or degraded antibody • Check manufacturer’s recommendations for storageand/orexpirationdate.Ifexpiredorpast storage time, purchase new antibody. • Ifantibodywassubjectedtorepeatedfreeze/thaw cycles purchase new antibody, as this can affect thestructureandprotein:antibodybinding.

Non-specific binding of primary •Decreaseconcentrationofprimaryantibody. antibody •Checkconcentrationofproteinbeingaddedtogel. •Useamorespecificantibody,suchasa monoclonalantibodyorAntigenAffinityPurified polyclonal antibody. •Addasmallamountofdetergent,suchas TWEEN-20 (0.1-0.5%) to the primary antibody solutionand/orblottingbuffer. • Increasenumber/durationofwashes.

Non-specific binding of secondary •Decreaseconcentrationofsecondaryantibody. antibody •Runacontrol,omittingtheprimaryantibody incubation.Ifbandsdevelop,chooseadifferent secondary antibody. •Addasmallamountofdetergent,suchas TWEEN-20 (0.1-0.5%) to the secondary antibody solution. • Increasenumber/durationofwashes.



Appendix IV

Immunohistochemistry

The analysis of tissues and cells for the presence of specific proteins is an important component of numerous clinical diagnos-tic and research programs. Immunohistochemistry (histo=tissue) and immunocytochemistry (cyto=cell) are powerful techniques that utilize the specific interaction between a target protein and its cor-responding antibody to visualize a protein of interest in a complex biological sample. The terms immunohistochemistry (IHC) and im-munocytochemistry are often used interchangeably due to the inti-mate relationship between cells and tissues.

Two common detection methods used in IHC are immuno-fluorescence (IF) and immunoenzymatic staining. In IF, the direct or indirect detecting antibody is labeled with a fluorophore, such as fluorescein or rhodamine. In immunoenzymatic staining, the direct or indirect detecting antibody is conjugated to a chromogen activat-ing enzyme such as horseradish-peroxidase (HRP) or alkaline phos-phatase (AP). Thus, there are both direct and indirect IHC protocols, where the researcher selects the best method of detection for the tis-sue source and antigen of interest. This flexibility creates a variety of IHC protocols, including an indirect polymer labeling method for non-biotin based systems which enhances the signal. Please note that an indirect immunoenzymatic detection method is illustrated on the next page (Figure IV-1).

Working with cells and tissue can pose difficult problems due to the elaborate and complex nature of these structures. In order to accurately examine these complex samples it is vital to preserve, to the extent possible, the native state of the tissue being analyzed. This is accomplished through a technique known as fixation. The fixation process is a cornerstone in immunohistochemistry and helps

maintain the integrity and authenticity of the proteins and structures present in the samples. This is usually achieved by inhibiting the innate degradative processes present in the sample tissue by some combination of cryostasis, treatment with chemical fixatives, and embedding tissue samples in paraffin. One of the most commonly used fixative combinations is the formalin-fixed paraffin-embedded (FFPE) fixation method. To achieve maximum recognition of the tar-get protein, it is necessary to optimize the fixation process for each sample type analyzed. Factors which influence the optimization of the fixation process include the purity, affinity and specificity of the detecting antibody, the structure and stability of the target protein, and the structural and biochemical characteristics of the tissue sam-ple.

The fixation process, while critical for maintaining the integ-rity of the tissue, often masks the antibody recognition epitope on the target protein by inducing chemical modification, conformational alterations, or protein aggregation. Therefore, most IHC procedures contain an unmasking step which is frequently called antigen retriev-al, or epitope recovery. This unmasking process can be accomplished by several procedures, including HIER (Heat-Induced Epitope Re-trieval) and PIER (Protease-Induced Epitope Retrieval), which, de-pending on the sample and the target protein, can be used alone or in tandem.

The detection strategies commonly applied in IHC procedures can be classified into two general categories, designated as direct and indirect detection methods. The direct detection methods utilize a labeled antibody, capable of fluorescing or activating a chemical chromogen, which binds directly to the target protein. This method


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has the advantages of simplicity, low non-specific binding in most cases, and short assay run times. Indirect methods use an unlabeled primary antibody to initially bind the target protein, which is then recognized by a labeled secondary antibody. Some indirect IHC de-tection procedures use a three stage procedure, in which an unlabeled secondary antibody is used and subsequently recognized by a labeled tertiary antibody. Most indirect methods produce some degree of sig-nal amplification during the multiple stages of the detection proce-dure, which consequently results in increased assay sensitivity.

Figure IV-1 is a sample IHC detection protocol representing the use of an indirect immunoenzymatic method.

Note: Researchers modify protocols in order to prevent nonspecific staining such as the blocking of endogenous proteins or enzymatic molecules.

References (1) Boenisch, Thomas ed., Immunochemical staining procedures

handbook. 3rd ed. Carpinteria, CA; 2001.(2) Janeway, et al., Immunobiology: the immune system in health

and disease. 6th ed. New York: Garland Science, 2005.

Figure IV-1 - Indirect Immunoenzymatic Detection Method

Immunohistochemistry Continued



Anti-Human CXCL-16(Catalog #500-P200)

Human colon/rectum adeno-carcinoma (FFPE) stained with Rabbit Anti-Human CXCL-16

at 0.5 µg/mL.Tissue samples were provided by the

Cooperative Human Tissue Network, which is funded by the National Cancer Institute.

Other investigators may have received speci-mens from the same subjects.

Anti-Human IL-6 (Catalog #500-P26)

BJ ras cells stained with Rabbit Anti-Human IL-6 at 3.0 µg/mL. Information and photo are courtesy of the Cell Profiling group, SciLifeLab Stockholm.

Anti-Murine RELMα (Catalog #500-P214)

Murine lung tissue (4% PFA from infected mice) stained with Rabbit Anti-Murine RELMα. Information and photos are courtesy of Meera Nair and David Artis, University of Pennsylvania.

Photographs, PEPROTECH, © 2011


Immunohistochemistry Troubleshooting Guide



Weak/No Staining Tissue Processing •Theantigenmayhavebeendestroyedduringthetissueprocessing. •Ensuretissueispreparedproperlypriortoimmunostaining procedure.Useproperfixative,fixationtime,embeddingreagents andembeddingtechniques.Usepropersectioningtechniques andequipment.TherecommendedtissuethicknessforIHCis 4µmsections.Forformalin-fixed,paraffin-embeddedtissue, ensureslidesarecompletelydeparaffinizedandrehydrated. Replace xylenes and alcohols as needed.

Antigen Retrieval •Determineoptimalantigenretrievalconditions.Considerstaining (Epitope Retrieval) atissuesectionwithoutantigenretrieval. •UseHIER(HeatInducedEpitopeRetrieval).Adjustandmaintain thetemperature.AdjustthepH(chooseappropriatebuffer).Adjust the amount of time. •UseEIER(EnzymeInducedEpitopeRetrieval).Choosean appropriateenzyme(e.g.proteinaseK).Adjusttheincubation time.Performantigenretrievalatroomtemperatureorwithheat. •Donotreuseantigenretrievalsolution.Performadequatewash steps.

Primary Antibody •Ensureantibodiesarepreparedasperdatasheet: •Centrifugevialpriortoopening. •Oncetheantibodyisreconstituted,centrifugevialtoremove any possible aggregates. •Aliquotandfreezeantibodyat-20˚C.Avoidrepeatedfreeze/ thaw cycles. •Useantibodybeforeexpirationdate.

•Increaseantibodyconcentration. •Increaseincubationtime(e.g.stainantibodyovernightat4˚C).

Secondary Antibody •Matchprimaryantibodywithappropriatesecondaryreagentor detection system. •Increasesecondaryantibodyconcentrationorincubationtime.

Detection System •Useappropriatedetectionsystem. •Ensureenzymeandchromogenarecompatible(e.g.DAB chromogenrequirestheHRPenzyme). •Usereagentsinproperorder. •Performappropriatewashsteps.

continued…




Weak/No Staining Detection System continued… continued continued… •Usecomponentsatroomtemperature. •Usecomponentspriortoexpirationdates. •Useanamplifieddetectionsystem,suchasapolymerdetection systemortyramideamplification.Adirectlabelingsystemmay havelessamplificationandmayproduceaweakersignal. •Sodiumazideisaninhibitorofhorseradishperoxidaseandmay interfere with staining results if present in certain reagents, such asanHRP-conjugatedsecondaryantibody.Limitedamountsof sodium azide in other reagents should not interfere with the immunostaining procedure.

Antigen •Thisantigenmaynotbepresentintheparticulartissueorata significantlevelfordetection. •Theepitopesoftheantigenmayhavebeendestroyedduring tissueprocessingorantigenretrieval.Also,ifusingamonoclonal antibody and that particular epitope is masked or destroyed, then the antibody will not stain. Consider re-staining or reprocessing a new tissue sample. •Useanalternativeassaytodetectforthepresenceoftheantigen (e.g. in situ hybridization).

Chromogen •Adjustincubationtime.Checkforchromogenprecipitateanddo not use past expiration date. •Useappropriatemountingmediaforeachchromogen(e.g.DAB canbecoverslippedwithapermanentmountingmedia,whilea non-alcoholsolubleAECwillrequireanaqueousmountingmedia).

Over-staining Tissue Processing •Ensuretissueispreparedproperlypriortoimmunostaining procedure.Useproperfixative,fixationtime,embeddingreagents andembeddingtechniques.Usepropersectioningtechniques andequipment.TherecommendedtissuethicknessforIHCis 4µmsections.Forformalin-fixed,paraffin-embeddedtissue, ensureslidesarecompletelydeparaffinizedandrehydrated. Replace xylenes and alcohols as needed.

Antigen Retrieval •Thetissuemaybeover-retrieved.Adjusttheincubationtime. Adjustthetemperature.Ifnecessary,usealternateantigen retrievalmethods.Stainatissuesectionwithoutantigenretrieval.

Primary Antibody •Ensureantibodiesarepreparedasperdatasheet: •Centrifugevialpriortoopening. •Oncetheantibodyisreconstituted,centrifugevialtoremoveany possible aggregates. continued…




Over-staining Primary Antibody •Aliquotandfreezeantibodyat-20˚C.Avoidrepeatedfreeze/thaw continued… continued… cycles. •Useantibodybeforeexpirationdate. •Decreaseantibodyconcentration.Adjustantibodyincubationtime. •Incubatetissuespecimenwithaproteinblockjustpriortothe primary antibody incubation (see protein block below).

Secondary Antibody •Adjustconcentrationofsecondaryantibody.Useasecondary antibodythatislesslikelytoexhibitnon-specificbinding(e.g. cross-adsorbed antibody or F(ab’)2 fragment).

Detection System •Useadetectionsystemthatwillexhibitlessnon-specificbinding. •Adjustincubationtime(s).

Non-specific Binding •Blockendogenousmoleculesandhighlychargedmolecules.

Endogenous •Dependingonthetissuetypeandstainingprocedure,endogenous Molecules molecules may be present and interfere with staining. Determine the necessary blocking steps for each tissue, such as blocking for endogenous molecules such as endogenous biotin, endogenous peroxidase, endogenous phosphatase, or Fc Receptors. •Chooseanalternativestainingmethodwhenanendogenous moleculemaybedifficulttocompletelyblock(suchasanon- biotindetectionmethodonliver,whichisrichinbiotin).

Protein Block •Useaproteinblockorserumblockfromthehostanimalofthe secondaryantibodytopreventnon-specificbinding(antibodies are highly charged proteins and the molecules’ hydrophobicity maycausenon-specificstaining). •Incubatethetissuespecimenwiththeproteinblockpriortothe primary antibody incubation.

Inadequate washing •Increasetheamountoftimeand/orthenumberofwashes.Use the appropriate buffer pH for different staining systems (e.g. PBS or Tris wash buffers). Add a detergent, such as TWEEN-20 to reduce hydrophobic interactions.

Immunohistochemistry Troubleshooting Guide Continued



Over-staining Chromogen •Adjustincubationtime.Ifpossible,adjustthechromogen continued… concentration. Check for precipitate and do not use past expiration date.

Tissue sections •Donotallowtissuesectionstodryoutduringtheimmunostaining dried out procedure.Useahumiditychamberandsufficientreagent volumeduringtissueincubations.UseaPAPpentocreatea hydrophobic barrier around the tissue section to ensure the tissue remainscoveredwithareagent.

Other Conditions Reagent Expiration •Checktheexpirationdatesofallreagentsusedduringthe Dates immunostaining procedure.

Controls •Ensurepositiveandnegativecontrolsarevalid.

Surfactant wash •Addasurfactantwashsteptoenhancereagentpenetration(e.g. PBS/0.2%TritonX100).

Mounting Medium •Useappropriatemediawitheachchromogen(different chromogensrequireaqueousornon-aqueousmountingmedias).

Counterstain •Counterstainfortheoptimalamountoftime.

Automated Staining •Checkmachinetoensureproperspecifications.



Appendix V

MicroarrayMicroarray technology enables the simultaneous detection

and quantification of a large number of proteins or nucleic acids in a variety of biological samples, including cells, tissues, and bodily fluids. The term microarray refers to a miniaturized solid support or chip, such as a nylon membrane or glass microscope slide, which is spotted with a capture element (antibody, protein, oligonucleotide, cDNA, etc.) in a grid-like configuration. There are many applications of microarrays including assessing protein families, probing serum for diagnostic information, profiling expression in response to drugs, and profiling cells treated with certain stimulators or transfected with a given gene. Although there are many procedural variations, the five essential steps in the preparation of a microarray are: 1) capture element synthesis, 2) preparation of the solid support surface, 3) im-mobilization of the capture elements onto the solid support (using a robotic arrayer), 4) binding of the target molecule to the immobilized capture elements, and 5) detection and quantification of the target/capture element complex.

Antibody microarrays use antibodies, raised against known protein antigens, as the capture elements and proteins capable of spe-cifically binding to the immobilized antibodies as the detected target molecules. Fluorescent dyes such as Cy3, Cy5, FITC, and Texas Red, or enzyme systems such as biotin-horseradish peroxidase, are a sensitive means of detection and visualization. Antibody microar-rays exploit many of the same principles applied to well established immunoassay procedures such as ELISA or Western blot; however, microarrays offer many advantages including an increased number of data points (which has a direct effect on accuracy), the ability to detect many proteins simultaneously, and reduced antibody and sample volume requirements. In addition, once an array has been

designed and the components validated, a robotic arrayer can rapidly spot limitless copies for use with large sample groups.

In this article, two antibody microarray formats will be highlighted: direct labeling and fluorescence-linked immunosorbent assay (FLISA). Both assays use specific antibodies as the capture element. In direct labeling, all proteins in the sample mixture are labeled prior to capture in contrast to FLISA, where target proteins from an untreated sample mixture are captured and subsequently de-tected through the use of a biotinylated antibody and a fluorophore-labeled secondary antibody.

Antibodies used in microarray assays work best when they are highly purified, exhibit minimal cross-reactivity, and have been shown to work well in traditional ELISA formats. Initially, antibody solutions are prepared at a pre-determined optimal concentration (usually 300-500µg/ml), and then they are assembled into a print plate or microtiter plate, which is used by a robotic arrayer to prepare a spotted solid support. Six to ten replicates per antibody are usually sufficient and, depending on the researcher’s needs, up to 500 differ-ent antibodies per array can be spotted and immobilized for compari-son. The spotted support is blocked by treatment with a BSA solution for at least one hour, and then the support is ready to be exposed to the samples of interest (serum, urine, cell lysate, etc).

Figure IV-1 depicts direct labeling detection. All proteins in a complex mixture are conjugated to either a fluorophore (Figure V-1A), or biotin, prior to incubation with the capture element. The arrays are then washed and scanned for data analysis. The compari-son of signal intensities between samples can be used to measure the relative amounts or expression levels of detected proteins. Al-ternatively, the comparison of signal intensities to calibrated protein


Appendix V continued

standards, labeled with distinct fluorphores, allows exact expression levels to be quantitatively measured (Figure V-1B). Advantages of this method are its simplicity and the requirement of only one specific antibody per target. The primary disadvantage is increased background signal resulting from the labeling of all proteins in the

sample mixture (e.g. albumin in serum). Detection sensitivity is lim-ited by the concentration of the background protein relative to the target protein and is typically around 100ng/ml in serum.

In FLISA detection (Figure V-2), antibodies spotted on the microarray capture unlabeled protein targets. This is followed by in-

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Figure V-1A - Direct Fluorophore Labeling

Figure V-1B - Multi-Color Detection


Appendix V continued

cubation with a cocktail of biotin-labeled antibodies, each antibody corresponding to one of the spotted capture antibodies. Finally, the array is incubated with a suitable detection antibody such as a fluoro-phore labeled anti-biotin secondary antibody. The FLISA detection format results in increased sensitivity, relative to the direct labeling method, because background is significantly reduced. Sensitivity in this format can be enhanced even further using tyramide signal amplification or rolling-circle amplification (3). However, optimi-zation of assays measuring many targets can be difficult using this format; therefore, FLISA detection is usually used for a limited num-ber of targets that are present at concentrations below the detection

limit of the direct labeling method. Although the detection limits for the assay depends on a number of parameters, including the affinity and specificity of the antibodies used, the protein background in the sample, and the detection conditions chosen, the FLISA method can provide detection limits in the low pg/ml range.

References:(1) Fung, Eric ed., Protein arrays: methods and protocols. Fremont, CA: Humana

Press Inc.; 2004.(2) Ye, S. and Day, I. eds. et al., Microarrays and Microplates: applications in

biomedical sciences. Oxford, UK: BIOS Scientific Publishers Ltd.; 2003.(3) Zhou, Heping, et al., Genome Biology, Vol. 5, issue 4, R28; 2004.

Figure V-2 - FLISA

Microarray Continued


Appendix VI

Flow Cytometry (FCM)Flow cytometry (FCM) is an extremely valuable research

tool for analyzing various microscopic particles, such as individ-ual cells. The development of this powerful technique represents the culmination of advances in several seemingly diverse scientific fields, including fluid mechanics, laser systems, optics, electronics, and computer programming. One of the major strengths of FCM is the capacity for multiparametric analysis of complex cell mixtures. FCM can be used to simultaneously determine multiple character-istics of individual cells within the mixed population, including the size, structural complexity, phenotype, and sub-type. Depending on the equipment being used, it is possible to analyze hundreds of indi-vidual samples per second.

The seemingly improbable task of evaluating multiple param-eters from samples containing such a vast and divergent number of cells is accomplished through the hydrodynamic focusing of the cells suspended in a suitable buffer, which allows for the optical and electrical systems’ individual analysis of each cell. Depending on the amount of information a researcher desires to collect from the samples, one or more lasers may be used for the analysis of the samples. Considering the complexity of orchestrating such a wide and varied range of systems, the accomplishment of such a feat re-lies heavily on the flow cytometer instrumentation.

An extended application of FCM technology is the ability to separate cells according to pre-determined characteristics deemed important by the researcher. Once they have been categorized us-ing these selected factors, an automated system collects the cells in designated receptacles for further study.

Fluidic System Fluid mechanics, or the study of the properties of fluids (in

this case, liquid) and their reactions to various forces, is one of the disciplines that must be utilized in FCM. In order to obtain reliable results from the analysis of samples while maintaining a high rate of analysis, it is crucial to ensure that the cells are passing through the observation point one at a time (Figure VI-1). Several factors must be considered in order to achieve the sample presentation required for this application. First, the cells must be completely suspended in a suitable solution; the presence of clusters or groupings will con-found the analysis and may clog the liquid handling components of the flow cytometer. Samples derived from blood, for the most part, already fit this requirement, while samples derived from whole tis-sue require a separate protocol in order to disassociate the cells from extracellular matrix components, and homogenize the sample.

Figure VI-1 - Hydrodynamic Focusing


Appendix VI continued

Flow Cytometry (FCM) Continued

Second, it is necessary to direct the cells, one at a time, through an intersection of the fluid stream and a laser beam. This is achieved through a process known as hydrodynamic focusing and occurs in a device referred to as the flow chamber. While the purpose of hy-drodynamic focusing can be simply stated as encasing the sample, or core fluid containing the cells, inside a sheath fluid resulting in “single file” presentation, obtaining this precarious state is anything but simple. The achievement of this spatial relationship utilizes a principle of fluid dynamics referred to as laminar flow. In laminar flow, the two fluid streams remain separate with no turbulence be-tween them allowing the cells to move in single file for analysis. A critical parameter in accomplishing this carefully balanced state is the relative pressures of the two liquid components. Maintain-ing the appropriate proportion of the pressure and flow rate for the sheath fluid in comparison to that of the core (cell containing) fluid is required in order to successfully execute the hydrodynamic focus-ing process. The higher pressure of the sheath fluid accelerates the core fluid, focusing the cells in the middle of the stream. This al-lows the laser to intersect the core stream and analyze the cells one at a time, and with maximum efficiency.

Optical and Electronic SystemsThe foundation for various modern assays relies on the analy-

sis of light, or more accurately, electromagnetic radiation. Advances in the development of critical components and highly specialized materials have enabled faster, more efficient optical analysis at re-duced costs, and have helped to propel FCM into a wider range of practical applications. There are several main components that com-prise the basic optical system of the flow cytometer (Figure VI-2).

The light source, which typically consists of a single laser, but can also consist of multiple lasers depending on the device in use and the parameters of the assay, acts as the excitation source. Filters and mirrors direct the beams and help differentiate the various wave-lengths of light. Finally, the detectors capture specified light signals and convert them to electrical signals.

There are two main categories of detection used in FCM: light scatter and fluorescence. In general terms, the information gener-ated by light scatter indicates the physical properties of the sample, whereas information generated from fluorescence relates more to biochemical properties, such as phenotype. In order to effectively detect light scatter and fluorescence, light detectors are placed at critical angles around the observation point. As the laser beam (in-cident beam) strikes a cell, the light of the beam will refract, or scatter, at various angles depending on certain characteristics of the cell.

The electronics present in the flow cytometer serve a basic but critical function in the analysis of the samples. As the laser beam interacts with the cells, the various detectors capture the scattered light and convert them into an electrical signal that can be inter-preted and processed by computer software. These detectors can be either photodiodes or photomultiplier tubes (PMT), each filling a specific role in the flow cytometer.

The light scatter detectors measure the intensity and corre-sponding refraction angles of the refracted beam, which falls into two main designations: forward scatter and side scatter. Forward scatter, or low angle scatter, is collected by a single detector known as the Forward Scatter Channel, or FSC, which is typically a pho-todiode. The FSC not only acts as a simple counter, but can also



Figure VI-2 - Fluorescence Flow Cytometer



Flow Cytometry (FCM) Continued

Labeled Antibody ReagentsA number of FCM applications utilize fluorophore conjugat-

ed antibodies to indicate the presence of a specific protein or other marker. These antibodies typically recognize specific epitopes, ei-ther on the cell’s surface or inside the cell itself. For most FCM applications, the best results are obtained when the conjugated an-tibodies being used are highly purified preparations that bind with high affinity and specificity to their target antigens. High affinity antigen binding usually leads to a stronger signal, resulting in in-creased sensitivity. Use of highly specific antibodies typically yields unambiguous results, minimally compromised by non-specific bind-ing and background signals. Accordingly, either affinity purified polyclonal antibodies and/or high affinity monoclonal antibodies are, in most cases, best suited for use in FCM. The characteristics of the fluorophore(s) used are also critical. As discussed above, the difference between the peak emission and excitation frequencies should be as large as possible. Additionally, when multiple fluoro-phores are used in a single run, it is necessary that each flourophore contain a distinct emission spectrum.

SoftwareConsidering the high number of complex components that

must function in concert with one another, it is no surprise to learn that computer software is an essential part of successful FCM opera-tion. The software serves several main functions that are necessary for the proper analysis of the vast number of test samples. One of the primary functions is to control all of the complex mechanical and temporal systems that allow the flow cytometer to operate ef-

detect characteristics of the sample from the refraction pattern. The characteristics revealed by forward scatter include the general size or volume, and the integrity of the cell. Side scatter, or wide angle scatter, is detected by one or more Side Scatter Channels (SSC). This form of scatter is generally more pronounced for cells with greater complexity since various structures, both internal and ex-ternal, tend to increase the amount of light scattering. Information gathered from side scatter can also be used to determine the general shape of the cell. Additionally, the SSCs can detect fluorescent sig-nals in assays that incorporate fluorescence.

The forward scatter and side scatter profiles of each cell type are unique, and when the information from each channel is com-bined, it is possible to accurately determine the cell types present in a heterogeneous population. More detailed information can be collected from the emission of added marker specific fluorophore conjugated antibodies.

Fluorescence is a form of luminescence that relies on the input of energy, in this case photons from the laser source, to increase the energy state of the molecule (fluorophore). The flourophore then emits light when reverting to a lower energy state. The excitation and emission characteristics (spectral properties) are important fac-tors in determining the suitability of flourophores to FCM applica-tions. Ideally, it is best to have as large of a difference as possible between the excitation and emission wavelengths (known as the Stokes Shift). If the excitation and emission spectra are too close to one another, or even overlap, it becomes increasingly difficult to obtain reliable results.



Figure VI-3 - Electrostatic Cell Sortingficiently; from the flow rates of the buffers, to the laser pulses ap-plied, to the detection and interpretation of the signals generated. The software is also responsible for creating and maintaining the files generated during the assay.

Once a run has been completed, the flow cytometry software functions to analyze the results. It is necessary to compile, interpret, and present a wide range of data coming from each sample. It is also important to differentiate between productive information and pos-sible contaminating or spurious data generated from the sample.

Fluorescence-Activated Cell Sorting (FACS™)Fluorescence-Activated Cell Sorting (FACS™*) is an extend-

ed application of flow cytometry. FACS™ can achieve the physical separation and collection of cells based on predetermined charac-teristics and parameters. During the FACS™ process, a specified fluorescence emission is detected, which triggers the impartation of a static charge by an integrated electrical charging ring. The charge is triggered just as the fluid stream is broken into droplets that typically contain one cell. The cell containing droplets then pass through an electromagnetic field where they are diverted based on charge, separated, and ultimately collected. The resulting isolated cell populations can then be further characterized or used in subse-quent research protocols.

*Becton, Dickinson and Company as the owner of the trademarked acronym FACS™


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