anti-phosphoserine/threonine Ab?

[T. S. Pillay]

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
and 
>they don't detect a lot of phosphorylated proteins.  Anti-pSer or pThr 
>antibodies are nowhere near as useful or specific as anti-pTyr
antibodies - 
>mores the pity.
>
>Jim
>
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".