Activation of PKC is one of the earliest events in a cascade leading to a variety of cellular responses such as secretion, gene expression, proliferation and muscle contraction (1,2).
PKC isoforms have been classified into three groups: classical PKCs, which are Ca2+ dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG) and phorbolesters (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 (3–7).
Members of these three PKC groups contain a pseudosubstrate 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.
PKCµ, 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 the control of three distinct phosphorylation events.
Specifically, Thr500 at the activation loop, the Thr641 autophosphorylation site and the Ser660 hydrophobic site at the carboxy-terminus of PKCb II are phosphorylated in vivo (2).
For the atypical PKC isoforms, there is no phosphorylation in the hydrophobic region, which has a glutamic acid residue in place of the serine or threonine residue found in other PKC isoforms.
The enzyme PDK1, or perhaps aclose relative, is responsible for PKC activation loop phosphorylation in a phosphoinositide 3 kinase (PI3K) dependent fashion (8,9).
Applications:Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000Immunohistochemistry: (Paraffin) 1:25IHC Protocol: Citrate/TBSTImmunoprecipitation: 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 immunoaffinity chromatography.
Form：Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM NaCl, 100ug/ml BSA, 50% glycerol.
Specificity：Detects endogenous levels of total PKCz. Does not crossreact with endogenous levels of other PKC isoforms.
Isotype：IgG
Calc Applications Abbrev：IHC IP WB
Calc Crossreactivity：Hu
Immunogen：Synthetic peptide (KLH-coupled) corresponding to human PKCz.
