Protein Kinase C is an 82kDa monomeric enzyme consisting of a C-terminal catalytic domain and a cysteine-rich N-terminal regulatory domain. The regulatory domain contains the sites for calcium and phospholipid binding and a pseudosubstrate subdomain, the target for PKC autophosphorylation. PKC is isolated from rat brain following the procedure of Walton and colleagues. The purified PKC consists primarily of α, β and γ isoforms with lesser amounts of δ and ζ isoforms.
Storage Buffer: 20mM Tris-HCl (pH 7.5), 2mM EGTA, 2mM EDTA, 1mM DTT, 10mM potassium phosphate (pH 7.5), 0.05% Triton® X-100 and 50% glycerol.
Unit Definition: One unit is the amount of kinase needed to transfer 1nmol of phosphate per minute using Type III-S histone protein as the substrate at 30°C in the presence of 0.6mg/ml phosphatidyl serine.
Walton, G.M. et al. (1987) Anal. Biochem. 161, 425–37.