Catalog # PP1091

PKCα (Ser-657/Tyr-658), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
ICC	1:100
IP	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Rb

MW 82 kDa

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.

References

Kawakami et al. (2003) PNAS. USA 100:9470-9475.
Nishizuka, Y. (1988) Nature 334:661.

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.

Immunocytochemical labeling of PKC phosphorylation in aldehyde-fixed and NP-40-permeabilized NGF-differentiated PC12 cells. The cells were labeled with rabbit polyclonal anti-PKCα (Ser-657/Tyr-658) (PP1091) antibody in the absence (Left) or presence (Right) of blocking peptide (PX1095). The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594.

Phospho-PKCα (Ser-657/Tyr-658) synthetic peptide corresponds to amino acid residues around serine 657 and tyrosine 658 of human PKCα. This sequence is similar to the conserved sites in rat and mouse, and has high homology to dual sites in human, rat, and mouse PKCβI (Ser-661/Tyr-662) and PKCγ (Thr-674/Tyr-675).

*For more information, see UniProt Accession P17252

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody detects an 82kDa* protein corresponding to the molecular mass of phosphorylated PKCα on SDS-PAGE immunoblots of neonatal rat brain lysate. Similar results were observed in human SKN-SH, endothelial, and HeLa cells, as well as rabbit spleen fibroblasts and rat pituitary cells. In immunoprecipitation experiments with various PKC isoforms, this antibody detected PKCα and PKCβ but not other PKC isoforms in rat brain lysate.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: