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 (1,2).
PKC isoforms have been classified in three groups: classical PKCs, whichare 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 (3-7).
These three PKC groups contain a pseudo-substrate orautoinhibitory 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 catalyticdomain.
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 C1domain 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 atthe activation loop, the threonine 641 autophosphorylation site and the serine 660 hydrophobic site at the carboxy terminus of PKCbeta•• 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 a close relative is responsible for PKC activation loop phosphorylation in a phosphoinositide 3 kinase (PI3K) dependent fashion (8).
Recommended Dilution: Western Blotting: 1:1000, incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight
Immunohistochemistry (Paraffin): 1:50
Optimal 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：rat
Grade：Affinity Purified
Purity：Purified by Protein A affinity chromatography.
Form：Supplied as liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.
Specificity：Detects endogenous levels of rat PKC delta only when phosphorylated at serines 643 and 676 of rat PKC theta. Does not crossreact with the phosphorylated PKC isoforms alpha, beta, gamma or epsilon. Species Crossreactivity: Human and monkey. Predicted to crossreact with mouse based on sequence homology.
Isotype：IgG
Calc Applications Abbrev：E IHC WB
Calc Crossreactivity：Hu Mk Mo Rt
Immunogen：Synthetic phosphopeptide (KLH coupled) corresponding to residues surrounding Ser643 of rat PKC delta.
