Phosphorylation of Proteins - Detection?

[Stephen R. Lasky]

In article <2vitui$q7q at dingo.cc.uq.oz.au>, forrest at biosci.uq.oz.au (acer) wrote:

> G'day all,
>         Does anyone know of a NON-radioactive procedure for testing whether a 
> protein is phosphorylated or not? 

You might try western blotting with antibodies against phosphotyrosine
(which I know are available) or phosphoserine or threonine (which I think
are now available).  Then detect with ECL.  We have done this with anti
P-Y antibodies and the technique is VERY sensitive.  To be even more
specific you could immunoprecipitate your protein with specific antibodies
before you run it on the gel for a western.  Hope that helps.

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Stephen R. Lasky, Ph.D.       Brown University/Roger Williams Medical Center
e-mail: Stephen_Lasky at brown.edu         LandLine: 401-456-6572
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America may be unique in being a country which has leapt from barbarism to decadence without touching civilization:  John O'Hara
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