Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

Phospho-PKC (pan) (ζ Thr410) (190D10) Rabbit mAb (Biotinylated) #5016

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M R Mk Endogenous 76-85 Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-PKC (pan) (ζ Thr410) (190D10) Rabbit mAb (Biotinylated) detects endogenous levels of PKC α, β I, β II, γ, δ, ε, η, θ and ι isoforms only when phosphorylated at a residue homologous to Thr410 of human PKCζ.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr410 of human PKC ζ.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or λ phosphatase-treated, using Phospho-PKC (pan) (ζ Thr410) (190D10) Rabbit mAb (Biotinylated) and developed with Streptavidin-HRP #3999.

Description

This Cell Signaling Technology (CST) antibody is conjugated to biotin under optimal conditions. The unconjugated Phospho-PKC (pan) (ζ Thr410) (190D10) Rabbit mAb #2060 reacts with human, mouse, rat and monkey phospho-PKC. CST expects that Phospho-PKC (pan) (ζ Thr410) (190D10) Rabbit mAb (Biotinylated) will also recognize phospho-PKC in these species.

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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Companion Products

Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.

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

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