Western blot analysis of extracts from various cell lines using PKCδ (D10E2) Rabbit mAb #9616.
Flow cytometric analysis of HeLa cells using PKCα Antibody (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Western blot analysis of extracts of HeLa, COS, C6 and NIH/3T3 cells, using PKCα Antibody.
Western blot analysis of extracts from 293, NBT-II, PC12 and COS cells, using PKCζ (C24E6) Rabbit mAb.
Western blot analysis of extracts from various cell lines using PKD/PKCμ (D4J1N) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of HeLa cell lysate and HeLa PKCδ KO lysate with PKCδ (D10E2) Rabbit mAb and β-Actin (D6A8) Rabbit mAb #8457.
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.
Confocal immunofluorescent images of C6 cells serum-starved (left) or TPA #9905 treated (center), labeled with PKCα Antibody (green) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Western blot analysis of extracts of Baculovirus expressed PKC isoforms demonstrating the isoform-specificity of PKCα Antibody.
Western blot analysis of bacterially expressed, GST-tagged, purified PKC isoforms, using PKCδ (D10E2) Rabbit mAb (upper) or GST (91G1) Rabbit mAb #2625 (lower), demonstrating specificity for PKCδ.
Western blot analysis of extracts from various cell lines using PKCδ (D10E2) Rabbit mAb.
|PKCα Antibody 2056||20 µl||
||H Mk M R||80||Rabbit|
|PKCζ (C24E6) Rabbit mAb 9368||20 µl||
||H Mk M R||78||Rabbit IgG|
|PKD/PKCμ (D4J1N) Rabbit mAb 90039||20 µl||
||H Mk||115||Rabbit IgG|
|PKCδ (D10E2) Rabbit mAb 9616||20 µl||
||H Mk M R||78||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
PKC Antibody Sampler Kit contains reagents to examine the total protein levels of various PKC isoforms. The kit contains enough primary and secondary antibodies to perform two Western blots per primary antibody.
All antibodies in this kit detect endogenous levels of total protein from their respective targets. All antibodies only recognize their specified isoform and do not cross-react with other PKC isoforms.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to human PKCζ and to residues surrounding Arg216 of human PKCδ protein and near the carboxy terminus of human PKD (PKCμ). Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the sequence of the human protein PKCα. 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).
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