Am 03.03.2010, 08:19 Uhr, schrieb Priscila Peña Diaz <[email protected]>:
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.
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