Product Pathways - Ca / cAMP / Lipid Signaling
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|
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.
Western blot analysis of extracts from NIH3T3 cells (lanes 1 & 2) and HeLa cells (lanes 3 & 4), untreated (lanes 1 & 3), PGDF-treated (lane 2), and TNFα-treated (lane 4), using Phospho- PKCζ/λ (Thr410/403) Antibody #9378, and PKCζ Antibody #9368.
Western blot analysis of extracts from 293 cells, untreated (lane 1), TPA-treated (.2μM for 20 minutes) (lanes 2 & 3), and CIP/λ phosphatase treated (lane 3), using Phospho-PKC (pan) (βII Ser660) Antibody #9371, Phospho-PKCα/βII (Thr638/641) Antibody #9375, and PKCα Antibody #2056.
Western blot analysis of extracts from C6 cells (lanes 1 & 2) and COS cells (lanes 3 & 4), untreated (lanes 1 & 3), or TPA-treated (.2μM for 20 minutes) (lanes 2 & 4), using Phospho-PKD/PKCμ (Ser916) Antibody #2051, Phospho-PKD/PKCμ (Ser744/748) Antibody #2054, PKD/PKCμ Antibody #2052, and β-actin Antibody #4967.
Western blot analysis of extracts from HeLa cells, untreated (lane 1), TPA-treated (0.2 μM for 20 minutes) (lanes 2 & 3), and CIP/λ phosphatase treated (lane 3), using Phospho- PKCδ (Thr505) Antibody #9374, PKCδ Antibody #2058, and β-actin Antibody #4967.
Western blot analysis of extracts from 293 cells, untreated (lane 1), TPA-treated (.2μM for 20 minutes) (lanes 2 & 3), and CIP/λ phosphatase treated (lane 3), using Phospho-PKCδ/θ (Ser643/676) Antibody #9376, Phospho-PKCθ (Thr538) Antibody #9377, PKCθ Antibody #2059, and β-actin Antibody #4967.
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.
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).
- Nishizuka, Y. (1984) Nature 308, 693-698.
- Keranen, L.M. et al. (1995) Curr. Biol. 5, 1394-1403.
- Mellor, H. and Parker, P.J. (1998) Biochem J. 332 (Pt 2), 281-292.
- Ron, D. and Kazanietz, M.G. (1999) FASEB J. 13, 1658-1676.
- Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Rep. 1, 399-403.
- Baron, C.L. and Malhotra, V. (2002) Science 295, 325-328.
- Flynn, P. et al. (2000) J. Biol. Chem. 275, 11064-11070.
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For Research Use Only. Not For Use In Diagnostic Procedures.