antibody

Protein Kinase C, delta, phosphorylated (Thr505) (PKCd)

P9103-19D

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

Protein Kinase C, delta, phosphorylated (Thr505) (PKCd)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

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). PKC isoforms have been classified in three groups: classical PKCs, which are Ca2+ dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG) and phorbol esters (TPA or PMA) through their cysteine-rich C1 domains; novel PKCs, which are Ca2+ independent but are still regulated by PS, DAG and TPA; and atypical PKCs, which are Ca2+ independent and do not require PS, DAG or TPA for their activation (3–7). These three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to the substrate binding site in the catalytic domain preventing its activation in the absence of cofactors or activators. Other members have been recently added to the PKC superfamily based on homology within the catalytic domain. PKCmu, or PKD, is regulated by DAG and TPA through its C1 domain. However, PKD is distinguished by a transmembrane domain and PH domain, as well as by its unique substrate recognition and Golgi localization. The PKC-related kinases, or PRKs, lack a C1 domain and thus do not respond to DAG or phorbol esters; instead they can be activated by phosphatidylinositol lipids, and their kinase activity is directly regulated by small GTPases of the Rho family through Rho binding to the homology region 1 (HR1). The activity of PKC is under control by three distinct phosphorylation events. Specifically, threonine 500 at the activation loop, the threonine 641 autophosphorylation site and the serine 660 hydrophobic site at the carboxy-terminus of PKCbeta II are phosphorylated in vivo (2). For the atypical PKC isoforms, there is no phosphorylation in the hydrophobic region, which has a glutamic acid 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 (8). Applications: Suitable for use in ELISA and Western Blot. Other applications not tested. Recommended Dilution: Western Blot: 1:1000, incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°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.

Hm Hu Mk Mo Rt

Synthetic phosphopeptide ccorresponding to residues around Thr505 of human PKC delta.

Recognizes endogenous levels of human PKC delta only when activated by phosphorylation at Thr505. Does not crossreact with the phosphorylated forms of PKC isoforms alpha, beta, gamma, zeta or epsilon. Species Crossreactivity: hamster, monkey, mouse and rat.

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.

Not determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not determined. Form: Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic phosphopeptide ccorresponding to residues around Thr505 of human PKC delta. Specificity: Recognizes endogenous levels of human PKC delta only when activated by phosphorylation at Thr505. Does not crossreact with the phosphorylated forms of PKC isoforms alpha, beta, gamma, zeta or epsilon. Species Crossreactivity: hamster, monkey, mouse and rat. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

E WB

-20°C

1. Nishizuka, Y. (1984) Nature 308, 693–698. 2. Keranen, L.M. (1995) Curr. Biol. 5, 1394–1403. 3. Newton, A.C. (1995) J. Biol. Chem. 270, 28495–28498. 4. Mellor, H. and Perker, P.J. (1998) Biochem. J. 332, 281–292. 5. Ron, D. and Kazanietz, M.G. (1999) FASEB J. 13, 1658–1676. 6. Way, K. J. et al. (2000) Trends Pharmacol. Sci. 21, 181–187. 7. Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Reports 1, 399–403. 8. Good, J.A. (1998) Science 281, 2042–2045. 9. Konishi, H et. al. (2001) Proc. Natl. Acad. USA 98, 6687–6592.

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