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9921
Phospho-PKC Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Phospho-PKC Antibody Sampler Kit #9921

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Phospho-PKC Antibody Sampler Kit: 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).

Phospho-PKC Antibody Sampler Kit: 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).

Phospho-PKC Antibody Sampler Kit: 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).

Phospho-PKC Antibody Sampler Kit: 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).

Phospho-PKC Antibody Sampler Kit: Image 5

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

Phospho-PKC Antibody Sampler Kit: Image 6

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

Phospho-PKC Antibody Sampler Kit: Image 7

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).

Phospho-PKC Antibody Sampler Kit: Image 8

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.

Phospho-PKC Antibody Sampler Kit: Image 9

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

Phospho-PKC Antibody Sampler Kit: Image 10

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).

Phospho-PKC Antibody Sampler Kit: Image 11

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).

Phospho-PKC Antibody Sampler Kit: Image 12

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

Phospho-PKC Antibody Sampler Kit: Image 13

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.

Phospho-PKC Antibody Sampler Kit: Image 14

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

Phospho-PKC Antibody Sampler Kit: Image 15

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.

Phospho-PKC Antibody Sampler Kit: Image 16

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

Phospho-PKC Antibody Sampler Kit: Image 17

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).

To Purchase # 9921T
Product # Size Price
9921T
1 Kit  (9 x 20 µl) $ 609

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

Product 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 two Western blot experiments.

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 (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.

Source / Purification

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.

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, 693-8.
  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.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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