Activation of PKC is one of the earliest events in the cascade leading to a variety of cellular responses such as secretion, gene expression, proliferation and muscle contraction.
PKC isoforms have been classified in three groups: classical PKCs, which are Ca2+ dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG) and phorbol esters (TPA or PMA) through their cysteine-rich C1 domains; novel PKCs, which are Ca2+ independent but are still regulated by PS, DAG and TPA; and atypical PKCs, which are Ca2+ independent and do not require PS, DAG or TPA for their activation.
These three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to the substrate binding site in the catalytic domain preventing its activation in the absence of cofactors or activators.
Other members have been recently added to the PKC superfamily based on homology within the catalytic domain.
PKCmu, or PKD, is regulated by DAG and TPA through its C1 domain.
However, PKD is distinguished by a transmembrane domain and PH domain, as well as by its unique substrate recognition and Golgi localization.
The PKC-related kinases, or PRKs, lack a C1 domain and thus do not respond to DAG or phorbol esters; instead they can be activated by phosphatidylinositol lipids, and their kinase activity is directly regulated by small GTPases of the Rho family through Rho binding to the homology region 1 (HR1).
The activity of PKC is under control by three distinct phosphorylation events.
Specifically, threonine 500 at the activation loop, the threonine 641 autophosphorylation site and the serine 660 hydrophobic site at the c-terminus of PKCbeta II are phosphorylated in vivo.
The enzyme PDK-1 phosphorylates and activates PKC delta at Thr505 in a phosphoinositide 3 kinase (PI3K) dependent fashion.
Applications:Suitable for use in ELISA, Western Blot and Immunoprecipitation.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000 Immunoprecipitation: 1:50Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage, store at -20°C.
Aliquots are stable for at least 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A and peptide affinity chromatography.
Form：Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.
Specificity：Recognizes endogenous levels of human PKC alpha, beta I, beta II, delta, epsilon and eta isoforms only when phosphorylated at a carboxy-terminal residue homologous to Ser660 of PKC beta II. Does not detect nonphosphorylated PKC or PKC phosphorylated at other sites. Apparent Molecular Weight: 78/80/82/85kD. Species Crossreactivity: mouse, rat and monkey.
Isotype：IgG
Calc Applications Abbrev：E IP WB
Calc Crossreactivity：Hu Mk Mo Rt
Immunogen：Synthetic phosphopeptide corresponding to residues surrounding Ser660 of human PKC beta II.
