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Technical Data Sheet

PK6540

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PKC (α,β,γ) Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The PKC (α,β,γ) phospho-regulation kit can be used to examine phosphorylation of PKCα (Ser-657/Tyr-658) relative to the total expression of PKCα. The phospho-specific antibody also detects phosphorylation of the the conserved dual site in PKCβ and PKCγ. The kit includes mouse monoclonal antibodies to detect total PKCα and PKC (α,β,γ), as well as secondary reagents for rabbit polyclonal and mouse monoclonal antibody detection.
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Western blot analysis of immunoprecipitates from neonatal rat brain lysate using anti-PKCα antibody. Control and alkaline phosphatase treated precipitates were probed with anti-PKCα (Central region) or anti-phospho-PKCα (Ser-657/Tyr-658). The latter shows no detection of PKCα after phosphatase treatment.

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Immunocytochemical labeling of PKC relative to F-actin in chick DRG neurons. The cells were labeled with mouse monoclonal PKC(α,β,γ) antibody (PM1101), then detected using appropriate secondary antibody (Red). This labeling is compared to F-actin staining (Green). (Image provided by Dr. Gianluca Gallo at Drexel University).

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Background
The Protein Kinase C (PKC) family of homologous serine/threonine protein kinases is involved in a number of processes such as growth, differentiation, and cytokine secretion. At least eleven isozymes have been described. PKC consists of a single polypeptide chain containing four conserved regions (C) and five variable regions (V). The N-terminal half interacts with PKC activators Ca2+, phospholipid, diacylglycerol, or phorbol ester, while the C-terminal half contains the catalytic domain. The conventional PKC subfamily (α, β1, βII, and γ) is regulated by both Ca2+ and diacylglycerol. The PKC pathway represents a major signal transduction system that is activated following ligand-stimulation of transmembrane receptors by hormones, neurotransmitters and growth factors. The phosphorylation of multiple sites in conventional PKCs regulates their activity. In mast cells, FceRI stimulation leads to phosphorylation of tyrosine 658 and 662 of PKCα and PKCβI respectively. This phosphorylation requires autophosphorylation of serine 657 and 661 in these respective kinases.

Background References
Nishizuka, Y. (1988) Nature 334:661.
Thiels, E. et al. (2000) J Neurosci. 20(19):7199.
Kawakami et al. (2003) PNAS. USA 100:9470-9475.
Buffer and Storage
Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.
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