anti-phospho S, T & Y antibodies

[Dr Engelbert Buxbaum]

Am 03.03.2010, 08:19 Uhr, schrieb Priscila Peña Diaz <ofiura from gmail.com>:

> Dear All:
>
> I've been looking recently into phosphorylation of certain membrane  
> proteins
> after a peptide antibody (specific for my protein of interest) fails to
> recognize the protein without prior treatment of the membrane (blotted
> proteins) with phosphatase.
>
> Most of the phosphorylation assays are usually performed with labelled
> phosphate (32P) but there seem to be antibodies that can also do the job  
> .
> I was wondering if anybody has experience with these monoclonal  
> anti-phospho
> S, T & Y antibodies. They look pretty fancy (and expensive) but would  
> like
> to hear if anybody has had any actual experience with them.

You can isolate phosphoproteins quite specifically by IMAC with Fe3+ or by  
titanium dioxide columns. MS/MS with electron capture dissociation is a  
good way to characterise the resulting proteins.