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Re: kinases protein kinase antibodies

Steven Pelech wrote:

> I am once again appealing for help from the protein kinase research
> community. Some of you may recall from my previous e-mail message two
> months ago to you that I have been interesting in identifying protein
> kinases that band shift on SDS-PAGE gels when they are phosphorylated. That
> list has not changed much since then.
> Currently, I am trying to identify protein kinase-specific antibodies that
> are potent and immunoblot fairly cleanly. The commercial distributors of
> signal transduction antibodies have not been very forth coming with samples
> of their antibodies for testing, and I can't afford to purchase all of the
> available antibodies that are sold to establish which ones really work. Of
> the forty or so commercial antibodies that I have tested, about a third
> that are sold do not recognize their target protein kinases when examined
> against a panel of 20 different rat and human cell lines and tissues. This
> is very disconcerting, but in fairness to the antibody supply companies, it
> can be very difficult to establish the specificity of an antibody when very
> little is known about its protein kinase target.
> I would greatly appreciate if everyone out there that uses antibodies to
> probe protein kinases could reply to this message by telling me which
> antibodies (i.e. the kinase antibody, the distributor and the catalogue
> number) you would recommend based on your experience. If there are
> antibodies that are really poor, this would also be valuable (at least I
> would be able to avoid them).
> If there are protein kinases for which antibodies are not currently
> available that you would like to see, please let me know. If you have large
> quantities of your own antibodies that you might be willing to trade or
> sell, I would also be very interested.
> I will provide the results of my findings about the performances of protein
> kinases antibodies with those that share their experiences with me.
> My long term objective is to develop a multi-blotting profiling service for
> the academic research community that can track the expression and
> phosphorylation states of over one hundred protein kinases at once. It may
> be possible to offer this service for under $2000. to track these kinases
> in a control and experimental sample. I believe that this approach could
> permit researchers to home in on those protein kinases that may be playing
> a significant role in their model system of interest. This is opposed to
> the current method, which is to go for the trendy kinases and hope for the
> best.
> Thank you 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

Dear Prof. Steven Pelech

I would like just to tell you about my personal experience of kinases
I used:
anti-PI3-K from UBI (# 06-195) good for IP less for WT
anti-src p60 from Santa-Cruz (# sc-N19) good for IP
anti-ACTIVE MAPK/JNK/p38 from Promega (# V6671/V7932/V2902) good for IP and WT
anti-ACTIVE MAPK from nanotools (# MAPK-12D4) good for IP.


Ouri Fischel, PhD
Synergix Ltd.
Technology Park
Malha, Building 1
91487 Jerusalem

E-mail: ouri@synergix.co.il
Fax: (972) (2) 6480399
Tel: (972) (2) 6480397