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RE: Protein kinase C activity assay? Help me!
You may want to use a better PKC substrate such as the short peptide named
"Kemptide". This peptide is a very good substrate, and it is suitable for
P-81 binding kinase assay.
Hope it will help, Yair
AstraZeneca R&D Charnwood
Discovery BioScience, Bakewell Road, Loughborough, Leics LE11 5RH,
Tel: +44 (0)1509 645481 Fax: +44 (0)1509 645557
From: awasthi [mailto:email@example.com]
Sent: 23 August 1999 18:19
Subject: Protein kinase C activity assay? Help me!
I have been doing radioactive PKC activity assay in crude extracts of
and cytosolic fractions of cells, using MBP 4-14 as substrate. I have been
getting high basal activity in the membrane fraction and that obscures the
induction of PKC activity by growth factors. The maximum increase I get is
doubling the basal activity. What may be the reason when literature is full
reports showing induction many-fold? Also, I get the induction only when
assay is done on membrane fraction and cytosolic fractions separately, why?
There are many more questions I will like to ask once I get response to
Details of the method may be important to answer, probably, which I will
provide on response.
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