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9921
Phospho-PKC Antibody Sampler Kit

Phospho-PKC Antibody Sampler Kit #9921

Western Blotting Image 1

Western blot analysis of extracts from NIH/3T3 cells, untreated, PDGF-treated (50 ng/ml) or TPA-treated (0.2 µM), using Phospho-PKD/PKCµ (Ser916) Antibody (upper) or PKD/PKCµ Antibody #2052 (lower).

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Western Blotting Image 2

Western blot analysis of extracts from various cell lines using PKD/PKCμ (D4J1N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Western Blotting Image 3

Western blot analysis of extracts from NIH/3T3 cells, untreated, PDGF-treated (50 ng/ml) or TPA-treated (0.2 µM), using Phospho-PKD/PKCµ (Ser744/748) Antibody (upper) or PKD/PKCµ Antibody #2052 (lower).

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Western Blotting Image 4

Western blot analysis of Baculovirus expressed PKCβ and PKCβ Ser660/Ala mutant, using Phospho-PKC (pan) (βII Ser660) Antibody (upper) or control PKCβ antibody (lower).

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Western Blotting Image 5

Western blot analysis of Baculovirus-expressed PKC isoforms alpha, beta, gamma, delta and epsilon, untreated or treated with lambda protein phosphatase, using Phospho-PKCalpha/beta II (Thr638/641) Antibody (upper) or PKCalpha, beta, gamma, delta, epsilon antibodies (lower).

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Western Blotting Image 6

Western blot analysis of extracts from U-937 cells, untreated or TPA treated (0.2 µM), using Phospho-PKCδ (Thr505) Antibody.

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Western Blotting Image 7

Western blot analysis of extracts from U-937 cells, untreated or TPA-treated (0.2 µM), using Phospho-PKCdelta/theta (Ser643/676) Antibody.

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Western Blotting Image 8

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-PKCθ (Thr538) Antibody #9377, PKCθ Antibody #2059, and β-actin Antibody #4967.

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Western Blotting Image 9

Western blot analysis of extracts from 293 cells, untreated or EGF-treated (100 ng/ml) and purified PKCzeta, untreated or lambda phosphatase-treated, using Phospho-PKCzeta/lambda (Thr410/403) Antibody (upper) or PKCzeta antibody (lower).

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Western Blotting Image 10

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 11

Western blot analysis of extracts from TPA, Go6983 and/or Bisindolylmaleimide treated 293 cells, using Phospho-PKC (pan) (βII Ser660) Antibody.

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Western Blotting Image 12

Western blot analysis of extracts from 293 cells, untreated, TPA treated (200 nM), or treated with TPA and CIP and λ phosphatases using Phospho-PKCα/βII (Thr638/641) Antibody or PKCα Antibody #2056.

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Western Blotting Image 13

Western blot analysis of extracts from Baculovirus-infected cells expressing PKC isoforms α, β, γ, δ or ε, using Phospho-PKCδ (Thr505) Antibody (upper) or control PKCα, β, γ, δ, ε antibodies (lower).

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Western Blotting Image 14

Western blot analysis of Baculovirus expressed PKC isoforms alpha, beta, gamma, delta and epsilon, untreated or lambda protein phosphatase-treated, using Phospho-PKCdelta/theta (Ser643/676) Antibody (upper) or PKCalpha, -beta, -gamma, -delta and -epsilon antibodies (lower).

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Western Blotting Image 15

Western blot analysis of Baculovirus expressed PKC isoforms, using Phospho-PKC (pan) (βII Ser660) Antibody.

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IHC-P (paraffin) Image 16

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-PKCdelta/theta (Ser643/676) Antibody.

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-PKCdelta/theta (Ser643/676). Antibody.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-PKD/PKCμ (Ser916) Antibody 2051 20 µl
  • WB
  • IP
H M R Mk 115 Rabbit 
PKD/PKCμ (D4J1N) Rabbit mAb 90039 20 µl
  • WB
  • IP
H 115 Rabbit IgG
Phospho-PKD/PKCμ (Ser744/748) Antibody 2054 20 µl
  • WB
H M R Mk 115 Rabbit 
Phospho-PKC (pan) (βII Ser660) Antibody 9371 20 µl
  • WB
H M R Mk 78, 80, 82, 85 Rabbit 
Phospho-PKCα/β II (Thr638/641) Antibody 9375 20 µl
  • WB
  • IP
H M Mk 80, 82 Rabbit 
Phospho-PKCδ (Thr505) Antibody 9374 20 µl
  • WB
H M 78 Rabbit 
Phospho-PKCδ/θ (Ser643/676) Antibody 9376 20 µl
  • WB
  • IHC
H M R Mk X 78 Rabbit 
Phospho-PKCθ (Thr538) Antibody 9377 20 µl
  • WB
H R Mk 79 Rabbit 
Phospho-PKCζ/λ (Thr410/403) Antibody 9378 20 µl
  • WB
H M R Mk 76 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two Western blot experiments.

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 (D4J1N) Rabbit mAb detects endogenous levels of PKD/PKCμ only. All other phospho-PKC antibodies recognize only their specific isoform when phosphorylated at the indicated sites.

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PKD. Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to the sequence of the human protein PKCβ II, PKCα, PKCδ, PKCθ, or PKCζ . 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).

  1. Nishizuka, Y. (1984) Nature 308, 693-8.
  2. Keranen, L.M. et al. (1995) Curr Biol 5, 1394-403.
  3. Mellor, H. and Parker, P.J. (1998) Biochem J 332 ( Pt 2), 281-92.
  4. Ron, D. and Kazanietz, M.G. (1999) FASEB J 13, 1658-76.
  5. Moscat, J. and Diaz-Meco, M.T. (2000) EMBO Rep 1, 399-403.
  6. Baron, C.L. and Malhotra, V. (2002) Science 295, 325-8.
  7. Flynn, P. et al. (2000) J Biol Chem 275, 11064-70.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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