Protein Kinase C, phosphorylated (PKC) BioAssay(TM) Sample Kit

Activation of PKC is one of the earliest events in the cascade leading to a variety of cellular responses such as secretion, gene expression, proliferation and muscle contraction (1,2). There are multiple PKC isoforms, including classical PKCs (a, b1, b2 and g), which bind calcium, diacylglycerol (DAG) and phospholipids; novel PKCs (d, e, n and theta), which are regulated by DAG and phospholipids; and atypical PKCs: zeta and lambda, which lack calcium-or DAGbinding domains. The activity of PKC is under control by three distinct phosphorylations. Specifically, threonine 500 at the activation loop, the threonine 641 autophosphorylation site and the serine 660 hydrophobic site at the carboxy terminus of PKCb2 are phosphorylated in vivo (2). For the atypical PKC isoforms, there is no phosphorylation in the hydrophobic region, which has a glutamic residue in place of the serine or threonine residue found in other PKC isoforms. The enzyme PDK1 or perhaps a close relative is responsible for PKC activation loop phosphorylation in a phosphoinositide 3 kinase (PI3K) dependent fashion (3). Protein Kinase D (PKD), also called PKCu, is a serine/ threonine kinase (4,5). Activation of PKD/PKCu is dependent on the phosphorylation of two activation loop sites, namely serine 744 and serine 748, via a PKCdependent signal transduction pathway (6). In addition to the two activation loop sites, the C-terminal serine 916 has been identified as an autophosphorylation site for PKD/PKCu. Phosphorylation of the serine 916 site is correlated with PKD/PKCu catalytic activity (7). Phospho-PKC Antibody Sampler Kit contains 40ul of each primary antibody and 100ul of antirabbit IgG secondary antibody (HRP conjugated).\n\nKit components: Crossreactivity: Application: Molecular Weight:\nP9103-22A: Protein Kinase C, pan, phosphorylated (PKC) 1x40ul\nrabbit H M R Mk W IP 78-82, 85 \nP9103-22B: Protein Kinase C, alpha, beta 2, phosphorylated (Thr638/641) (PKCa,bII) 1x40ul\nrabbit H M (R) Mk W IP 80, 82 \nP9103-22C: Protein Kinase C, delta, phosphorylated (Thr505) (PKCd) 1x40ul \nrabbit H M R Hm Mk W IHC 78 \nP9103-22D: Protein Kinase C, delta, phosphorylated (Ser643) (PKCd) 1x40ul \nrabbit H M R Mk W IHC 78 \nP9103-22E: Protein Kinase C, mu, Protein Kinase D, phosphorylated (Ser744/748) (PKCm, PKD) 1x40ul \nrabbit H M R Mk W IHC 115 \nP9103-22F: Protein Kinase C, mu, Protein Kinase D, phosphorylated (Ser916) (PKCm, PKD) 1x40ul rabbit H M R Mk W IP 115 \nP9103-22G: Protein Kinase C, mu, Protein Kinase D (PKCm, PKD) 1x40ul\nrabbit H M R Mk W 115 \nP9103-22H: Protein Kinase C, theta, phosphorylated (Thr538) (PKCt) 1x40ul \nrabbit H (M) R Mk W IHC 79 \nP9103-22J: Protein Kinase C, zeta, lambda, phosphorylated (Thr410/403) (PKCz,l) 1x40ul\nrabbit H M R Mk W IHC 76 \nP9103-22K: IgG, H&L (HRP) GtxRb 1x100ul \ngoat \n\nStorage and Stability:\nFor long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P9103-22

Size

9x40ul

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, BSA, 50% glycerol.

Purity

Purified by protein A and peptide affinity chromatography.

References

(1) Nishizuka, Y. (1984) Nature 308, 693-698. (2) Keranen, L.M. (1995) Curr. Biol. 5, 1394-1403.(3) Good, J.A. (1998) Science 281, 2042-2045. (4) Valverde, A.M. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8572-8576. (5) Johannes, F.J. et al. (1994) J. Biol. Chem. 269, 6140-6148. (6) Iglesias, T. et al. (1989) J. Biol. Chem. 273, 27662-27667. (7) Matthews, S. A. et al. (1999) J. Biol. Chem. 274, 26543-26549.

Additional Info

Detects PKCa, b1, b2, g, isoforms only when phosphorylated at a C-terminal residue homologous to Ser660 of PKCb2. Phospho-PKCd (Ser643) Antibody detects PKCd when phosphorylated at Ser643 and also detects PKCtheta when phosphorylated at Ser676. PKD/ PKCu detects PKD/PKCu only. All other phospho-PKC antibodies recognize only their specific PKCs when phosphorylated at the indicated sites.