Protein kinase C activity assay? Help me!

[awasthi <awasthi@uthscsa.edu>]

Dear kinasers,

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.

thanks

Awasthi
UTHSCSA, San antonio
210 567 4341

UTHSCSA, 7703 Floy Curl
San Antonio, TX 78284
Ph: 210 567 4341 (O)
    210 614 2637 (H)