anti-phospho S, T & Y antibodies

Priscila Peña Diaz via methods%40net.bio.net (by ofiura from gmail.com)
Thu Mar 4 06:01:18 EST 2010


On Wed, Mar 3, 2010 at 6:23 PM, Jayakumar, R <R.Jayakumar from roswellpark.org>wrote:

> We routinely do phosphotyrosine blots (human proteins) with
> anti-phosphotyrosine antibody clone 4G10 (many companies sell this).


Is it quite sensitive? Do your phosphorylated proteins bear several phospho
Y sites? Could you estimate how many cells/ug protein do you load for a nice
WB signal?


thanks


P.



>
> -----Original Message-----
> From: methods-bounces from oat.bio.indiana.edu [mailto:
> methods-bounces from oat.bio.indiana.edu] On Behalf Of Priscila Peña Diaz
> Sent: Wednesday, March 03, 2010 7:19 AM
> To: methods from magpie.bio.indiana.edu
> Subject: anti-phospho S, T & Y antibodies
>
> 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.
>
> kind regards
>
>
>
> Priscila
>
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