protein kinase C (PKC) inhibitors - (Sep/19/2007 )

Hi everyone,

I have to inhibited protein kinase C (PKC), the isoform alpha (PKC alpha) and the isoform 2 beta (PKC-2- simultanely in my cells (human myoblasts), and I have no idea wich PKC inhibitor to use. Also, I need an experiment to see if PKC is really inhibited.

Is anyone have worked with PKC inhibitor and can give my some advices?

Thanks!

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small molecular PKC inhibitors lack specificity; yet, in your case I recommend the indolocarbazole Gö6976 at recommended ~10 µM end concentration; Gö6976 is a cPKC inhibitor, which means you inhibit all cPKC alpha, betaI, beta II and gamma in parallel; more advanced is shRNA/siRNA technique

to check inihibition, you can do classical phosphotransferase activity tests utilizing radioactive ATP and a specific PKC peptide substrate or more advanced techniques with fluorescence substrates such as FRET (see f.i. Invitrogen) which change fluorescence after phosphorylating PKC-specific substrates;

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