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Re: kinases protein kinase antibodies



Dear Steven,

I used the following antibodies against protein kinases:

1. anti Bmx (Transduction Laboratories, B63620), WB (IP not tested)
2. anti Erk1 (Santa Cruz, sc-94), WB and IP (recognized Erk1 and Erk2 in both WB and IP)
3. anti Erk2 (Santa Cruz, sc-1647), WB and IP (specific for Erk2)
4. anti phospho-Erk (Santa Cruz, sc-7383), WB (IP not tested)
5. anti Jak2 (UBI, 06-255), WB and IP
6. anti Lyn (UBI, 06-207) WB (IP not tested)
7. anti p38 (New England Biolabs, 9212), WB (not for IP)
8. anti p38 (Santa Cruz, sc-7149), IP (not so good for WB)
9. anti phospho-p38 (New England Biolabs, 9211),  WB (IP not tested)
10. anti PI3-K p85 (Transduction Laboratories, P13020), WB (not for IP!)
11. anti Sam68 (Santa Cruz, sc-1238), does not work!
12. anti Syk (Santa Cruz, sc-573), WB and IP
13. anti Tec (Santa ruz, sc-1109), WB (IP not tested)
 

I used the following antibodies against protein phosphatases:

1. anti SHP-1 (UBI, 06-117), WB and IP
2. anti SHP-1 (Transduction Laboratories, P17320) WB and IP
3. anti SHP-2 (UBI, 06-118), WB (not for IP)
4. anti SHP-2 (Santa Cruz, sc-280), WB and IP
5. anti SHP-2 (Transduction Laboratories, P17420), WB and IP

WB = Westernblot, IP = Immunoprecipitation

Best regards,
Brigitte Kasper, PhD
Forschungszentrum Borstel
Abt. Immunologie und Zellbiologie
Parkallee 22a
23845 Borstel
Germany

phone: +49 4537 188 451
fax: +49 4537 188 404
e-mail: bkasper@fz-borstel.de
 

Steven Pelech schrieb:

Only half a dozen of the investigators who are part of this very large
protein kinase resource network answered my plea for the names of protein
kinase antibodies (and their suppliers and catalogue numbers) that they
have found either do or do not work. I know that you are all very busy out
there, but surely such information could be very helpful in your research
programs. I have personally found that about a third of the commercial
antibodies that I have tested DO NOT WORK OR BLOT VERY POORLY on the
kinases that they are meant to target. This results in a lot of wasted time
and money. I will provide a list of recommended and not recommended
antibodies from my personal findings and that of others to those that tell
me about their antibody experiences.

In view of the poor response to my question about protein kinase
antibodies, I am even more concerned about whether anyone is even
interested in using a multi-kinase blotting service for under $2000. where
about 100 different protein kinases could be screened by immunoblotting for
their expression and phosphorylation states in about 2 mg of crude cell
lysates from a control and an experimental sample (e.g. mitogen-, stress-
or drug-treated cells, or healthy and diseased tissue biopsies). You send
me the samples, I do the Western analysis and provide you with quantitative
information from densitometric scans of the immunoblots as well as copies
of the scans. This could allow you to focus on those kinases that are the
most markedly affected in your favourite model system. Most of you will
appreciate that the set up costs for such as analysis would be over
$25,000. to track about 100 kinases. I believe that this is much more
useful information than can be derived from gene expression arrays for
protein kinases.

If you think that such a multi-kinase blotting service would be useful,
please let me know (I will not follow up with further information about the
service in the future unless you request it in your e-mail reply). If you
don't think that such a service would be helpful for you, please also let
me know. A simple "YEA" or "NAY" e-mail reply for those of you who are
really busy would be most helpful.

Thank you in advance from Steven Pelech.

Steven Pelech, Ph.D.
Professor
Department of Medicine
Rm S125, 2nd Floor, Koerner Pavilion
2211 Wesbrook Mall
University of British Columbia
Vancouver, B.C.
Canada  V6T 1Z3

Office phone: (604) 822-8086
Office facsimile: (604) 822-8693
Confidential home facsimile: (604) 272-2102
E-mail: spelech@home.com