Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

PKCθ Antibody #2059

Applications Reactivity Sensitivity MW (kDa) Source
W IHC-P H M R Mk Endogenous 80 Rabbit

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

Specificity / Sensitivity

PKCθ Antibody detects endogenous levels of total PKCθ. The antibody does not crossreact with endogenous levels of other PKC isoforms.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) derived from the sequence of human PKCθ. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

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

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using PKCθ Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian adenocarcinoma, using PKCθ Antibody in the presence of control peptide (left), or antigen-specific peptide (right). Note specific staining of endothelial cells.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human basal cell carcinoma, showing cytoplasmic staining of tumor and endothelical cells, using PKCθ Antibody.

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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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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