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dephosphorylate a protein in cell lysate

Peter Cherepanov peter.cherepanov at uz.kuleuven.ac.be
Tue Jan 25 11:20:05 EST 2000


Hi people !

does someone know if it is possible to dephosphorylate a protein directly
in the cellular lysates ?  Or is it always necessary to immunoprecipitate
it first ?  And which phosphotase is the best to use for this purpose - SAP
or CIAP?
I tried 2 units of CIAP in NEB2 buffer for 10 mkg of total protein, but I
am nearly sure it is not enough.

(I just want to see a shift on my Western blot after dephosphorylation)

hope to get a hint,
Peter


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