Removal of free 32-P ATP

S. Tan sltan at u.washington.edu
Fri Jun 19 19:19:22 EST 1998


Hello:

I'm attempting to test whether my protein of interest is dephosphorylated
by a phosphatase.  I have immunopurified protein that can be
labeled by autophoshorylation in an in vitro kinase assay.  I would like
to use it as a substrate for the phosphatase in an in vitro
dephosphorylaton assay.  Should I be concern with the free ATP in the
kinase reaction solution?  It is OK to precipitate my labeled protein by
TCA precipitation and use the precipitate as a substrate for the
phosphatase assay?  Any advice and/or suggestion would be grealy
appreciated.

Stan




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