Analysis of the PKC-γ-Related Immunocrossreactive Region of a Novel Leukocyte Protein γ-rp

摘要: The newly discovered gamma-PKC-related-protein of human leukocytes (γ-rp) crossreacts with a polyclonal antibody preparation originally designed to be specific for PKC-γ (γMb-Ab). As this is currently the only suitable probe γ-rp, we sought characterize binding two proteins. We determined that γMg-Ab does not recognize native form γ-rp. However, denaturing immunoblots found 1) crossreactive γ-rp epitope differs somewhat from classic rat brain PKC-γ, but probably degree rat/human PKC species difference; 2) previously reported doublet bands represent single protein cell-stimulus inducible modifications; 3) antibodies present in pool bind separate sites within epitope; 4) access one site conformationally restricted, even after denaturation; 5) agonist-induced modification significantly affect total amount it can bind, 6) rate site. This investigation defines appropriate experimental use our antibody, and significance these findings future study cloning discussed.