Phosphorylation Assay Conditions

U12201 at uicvm.uic.edu U12201 at uicvm.uic.edu
Sat Jul 22 15:52:28 EST 1995


In article <1995Jul21.083008.11598 at leeds.ac.uk>, bmbrl at biovax.leeds.ac.uk says:
>
>I'm in the process of setting up an assay system to see if a membrane protein
>I've purified is a substrate for cAMP-dependent protein kinase (PKA).
>Firstly, what concentrations of ATP should I be using?  If I add [32P]ATP to
>the reaction mixture the final concentration will be ~0.1uM.  Would this be
>enough to indicate preferred substrates for phosphorylation?
>Any information would be gratefully received.
>Thanks, Rob.

I have had good results with the Pierce phosphocellulose units and the
kit for PKA (Prod # 29520 and 29530 respectively). The kit uses the
"kemptide" peptide substrate.
KeldS at uic.edu




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