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Phospho-PKC Antibody Sampler Kit #9921

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
Phospho-PKD/PKCμ (Ser916) Antibody #2051	40 µl	W IP	H M R Mk	115	Rabbit
PKD/PKCμ Antibody #2052	40 µl	W F	H M R Mk	115	Rabbit
Phospho-PKD/PKCμ (Ser744/748) Antibody #2054	40 µl	W	H M R Mk	115	Rabbit
Phospho-PKC (pan) (βII Ser660) Antibody #9371	40 µl	W	H M R Mk	78, 80, 82, 85	Rabbit
Phospho-PKCα/β II (Thr638/641) Antibody #9375	40 µl	W IP	H M Mk (R) (Hm)	80, 82	Rabbit
Phospho-PKCδ (Thr505) Antibody #9374	40 µl	W	H M R	78	Rabbit
Phospho-PKCδ/θ (Ser643/676) Antibody #9376	40 µl	W IHC-P	H M R Mk X	78	Rabbit
Phospho-PKCθ (Thr538) Antibody #9377	40 µl	W	H R Mk B (M)	79	Rabbit
Phospho-PKCζ/λ (Thr410/403) Antibody #9378	40 µl	W	H M R Mk	76	Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  X=Xenopus  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Phospho-PKC (pan) (βII Ser660) Antibody detects PKCα, β I, β II, δ, ε and η isoforms only when phosphorylated at a carboxy-terminal residue homologous to Ser660 of PKCβ II. Phospho-PKCδ (Ser643) Antibody detects PKC δ when phosphorylated at Ser643, and PKCθ when phosphorylated at Ser676. PKD/PKCμ antibody detects PKD/PKCμ only. All other phospho-PKC antibodies recognize only their specific isoform when phosphorylated at the indicated sites.

Description

The Phospho-PKC Antibody Sampler Kit provides a fast and economical means of evaluating multiple PKC isoforms and their phosphorylation state. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to the sequence of the human protein PKCβ II, PKCα, PKCδ, PKCθ, PKCζ or the mouse protein (PKD). 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 sites 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 occurs in vivo at Thr500 in the activation loop, at Thr641 through autophosphorylation, and at the carboxy-terminal hydrophobic site Ser660 (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. 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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Protocols

2051:

Immunoprecipitation, Western Blotting

2052:

Flow, Western Blotting

2054:

Western Blotting

9371:

Western Blotting

9374:

Western Blotting

9375:

Immunoprecipitation, Western Blotting

9376:

IHC / Paraffin, Western Blotting

9377:

Western Blotting

9378:

Western Blotting

Companion Products

• 2261 Phospho-(Ser) PKC Substrate Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9379 Phospho-PKC (pan) (gamma Thr514) Antibody
• 2056 PKCα Antibody
• 2058 PKCδ Antibody

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For Research Use Only. Not For Use In Diagnostic Procedures.