Phosphotyrosine antibodies

Glenn Cantor GCANTOR at VETMED.WSU.EDU
Tue Nov 7 14:57:42 EST 1995


------- Forwarded Message Follows -------

Date sent:      Mon, 06 Nov 1995 18:12:31 -0800
To:             gcantor at vetmed.wsu.edu
From:           mark ware <mark at terryfox.ubc.ca>

Dear Glenn Cantor,

        I noticed your inquiry regarding anti-phosphotyrosine antibodies. In
our lab we frequently use these antibodies to study cytokine receptor
signalling and we have tried all of the antibodies on the market. In our
hands we have found 4G10 to be superior to all other antibodies including
PY20 (which is probably why it is so much cheaper). For western blot
solutions we find 0.5ug/ml of TBS-Tween 20 to be sufficient for detecting
even low phosphorylation levels and this solution can be reused as many as
5-10 times with little loss of signal. You must be careful, however, to
block your membrane with 5% BSA and NOT skim milk powder, as skim milk has
phosphorylated proteins that will bind to your 4G10.

        If you need more information feel free to write me or you can look
at some of the papers from my lab. They can be found in an author search of
the names J. Damen, L. Liu, R. Cutler, and G. Krystal. Almost any of these
papers from the last 2-3 years will contain 4G10 immuno-precipitations and
westerns.   

Mark Ware



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