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kinases protein kinase antibodies



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