Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

PKC Isoform Antibody Sampler Kit #9960

Kit Includes Quantity Applications Reactivity MW (kDa) Source
PKCα Antibody # 2056 40 microliters W IP IF-IC F H M R Mk (Dg) 80 Rabbit
PKCδ Antibody # 2058 40 microliters W IP H M R Mk 78 Rabbit
PKCζ (C24E6) Rabbit mAb # 9368 40 microliters W H M R Mk 78 Rabbit
PKD/PKCμ Antibody # 2052 40 microliters W F H M R Mk 115 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dg=Dog

Specificity / Sensitivity

All antibodies in this kit detect endogenous levels of total protein from their respective targets. All antibodies only recognize their specified isoform and do not cross-react with other PKC isoforms.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated, PDGF-treated (50 ng/ml) or TPA-treated (0.2 µM), using PKD/PKCµ Antibody #2052.

Western Blotting

Western Blotting

Western blot analysis of extracts of HeLa, COS, C6 and NIH/3T3 cells using PKCα Antibody #2056.

Western Blotting

Western Blotting

Western blot analysis of extracts of HeLa, COS, C6 and NIH/3T3 cells using PKCδ Antibody #2058.


Western Blotting

Western Blotting

Western blot analysis of extracts from 293, NBT-II, PC12 and COS cells using PKCζ (C24E6) Rabbit mAb #9368.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to human PKCζ.Polyclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) derived from the sequence of the human protein PKCα, PKCδ, and mouse protein PKD (PKCμ). Antibodies are purified by protein A and peptide affinity chromatography.

Background

Activation of protein kinase C (PKC) is one of the earliest events in a cascade that controls a variety of cellular responses, including secretion, gene expression, proliferation and muscle contraction (1,2). PKC isoforms belong to three groups based on calcium dependency and activators. Classical PKCs are calcium-dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG) and phorbol esters (TPA, PMA) through their cysteine-rich C1 domains. Both novel and atypical PKCs are calcium-independent, but only novel PKCs are activated by PS, DAG and phorbol esters (3-5). Members of these three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to substrate-binding site in the catalytic domain to prevent activation in the absence of cofactors or activators.Control of PKC activity is regulated through three distinct phosphorylation events. Phosphorylation of Thr500 in the activation loop, the autophosphorylation site at Thr641 and at carboxy-terminal hydrophobic site Ser660 occurs in vivo (2). Atypical PKC isoforms lack hydrophobic region phosphorylation, which correlates with the presence of glutamic acid rather than the serine or threonine residues found in more typical PKC isoforms. Either the enzyme PDK1 or a close relative is responsible for PKC activation.A recent addition to the PKC superfamily is PKCμ (PKD), which is regulated by DAG and TPA through its C1 domain. PKD is distinguished by the presence of a PH domain and by its unique substrate recognition and Golgi localization (6). PKC-related kinases (PRK) lack the C1 domain and do not respond to DAG or phorbol esters. Phosphatidylinositol lipids activate PRKs and small Rho-family GTPases bind to the homology region 1 (HR1) to regulate PRK kinase activity (7).

  1. Nishizuka, Y. (1984) Nature 308, 693-698.
  2. Keranen, L.M. et al. (1995) Curr. Biol. 5, 1394-1403.
  3. Mellor, H. and Parker, P.J. (1998) Biochem J. 332 (Pt 2), 281-292.
  4. Ron, D. and Kazanietz, M.G. (1999) FASEB J. 13, 1658-1676.
  5. Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Rep. 1, 399-403.
  6. Baron, C.L. and Malhotra, V. (2002) Science 295, 325-328.
  7. Flynn, P. et al. (2000) J. Biol. Chem. 275, 11064-11070.

Application References

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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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