T. S. Pillay
tpillay at ucsd.edu
Thu Oct 20 10:21:23 EST 1994
Subject: Re: anti-phosphoserine/threonine Ab?
From: Jim Woodgett, jwoodget at oci.utoronto.ca
Date: Tue, 18 Oct 1994 15:00:39 GMT
In article <9410181000.AA39782 at bono.oci.utoronto.ca> Jim Woodgett,
jwoodget at oci.utoronto.ca writes:
>In article <Pine.3.89C.9410180802.A16076-0100000 at ciao.cc.columbia.edu>,
>js190 at COLUMBIA.EDU (Jae-Won Soh) writes:
>> Hi netters,
>> Is there any anti-phosphoSer/Thr antibody available on the market?
>Try Sigma. However, I don't recommend them as their specificity is lousy
>they don't detect a lot of phosphorylated proteins. Anti-pSer or pThr
>antibodies are nowhere near as useful or specific as anti-pTyr
>mores the pity.
I've done extensive displacement studies with antiPS/PT antibodies using
ELISA and I fully agree with Jim Woodgett- I've compared phosphate,
PS,PT,PY. PY and phosphate are highly effective at displacing these
antibodies-they lack specificity. There are a couple of groups who have
reported using these to study phosphorylation states but it if you speak
with them you'll get the same feedback-they don't work too well. If you
absolutely want to use antibodies, the best way is to make antibodies
against a phosphopeptide and use this as a probe. We have successfully
identified the major PKC phosphorylation site on the insulin receptor
using such approach- we raised an antibody to a synthetic serine
phosphopeptide - our paper will appear any day now in the Biochemical
Journal- "authors: Coghlan, Pillay, Tavare and Siddle".
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