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Protein kinase C activity assay? Help me!
I have been doing radioactive PKC activity assay in crude extracts of membrane
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 of
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 these.
Details of the method may be important to answer, probably, which I will
provide on response.
UTHSCSA, San antonio
210 567 4341
UTHSCSA, 7703 Floy Curl
San Antonio, TX 78284
Ph: 210 567 4341 (O)
210 614 2637 (H)