Description
Product Usage Information
Western Blotting: 1:1000
Immunoprecipitation: 1:100
Immunohistochemistry (Paraffin): 1:50
Immunofluorescence (Immunocytochemistry): 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized): 1:200 - 1:800
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 86952 .
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized)
Specificity / Sensitivity
PKC theta (E1I7Y) Rabbit Monoclonal Antibody recognizes endogenous levels of total PKCθ protein.
Species Reactivity: Human, Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro632 of human PKCθ protein.
Background
Activation of protein kinase C (PKC) is one of the earliest events in a cascade that controls a variety of cellular responses, including secretion, gene expression, proliferation, and muscle contraction (1,2). PKC isoforms belong to three groups based on calcium dependency and activators. Classical PKCs are calcium-dependent via their C2 domains and are activated by phosphatidylserine (PS), diacylglycerol (DAG), and phorbol esters (TPA, PMA) through their cysteine-rich C1 domains. Both novel and atypical PKCs are calcium-independent, but only novel PKCs are activated by PS, DAG, and phorbol esters (3-5). Members of these three PKC groups contain a pseudo-substrate or autoinhibitory domain that binds to substrate-binding sites in the catalytic domain to prevent activation in the absence of cofactors or activators. Control of PKC activity is regulated through three distinct phosphorylation events. Phosphorylation occurs at Thr500 in the activation loop, at Thr641 through autophosphorylation, and at the carboxy-terminal hydrophobic site Ser660 (2). Atypical PKC isoforms lack hydrophobic region phosphorylation, which correlates with the presence of glutamic acid rather than the serine or threonine residues found in more typical PKC isoforms. The enzyme PDK1 or a close relative is responsible for PKC activation. A recent addition to the PKC superfamily is PKCμ (PKD), which is regulated by DAG and TPA through its C1 domain. PKD is distinguished by the presence of a PH domain and by its unique substrate recognition and Golgi localization (6). PKC-related kinases (PRK) lack the C1 domain and do not respond to DAG or phorbol esters. Phosphatidylinositol lipids activate PRKs, and small Rho-family GTPases bind to the homology region 1 (HR1) to regulate PRK kinase activity (7). PKCθ is a novel protein kinase C that is predominantly expressed in T cells (8). Recruitment of PKCθ to the immunological synapse following T cell receptor stimulation plays an important role in the activation and proliferation of conventional T cells (9). Conversely, PKCθ negatively regulates the suppressive function of regulatory T cells and is excluded from regulatory T cell immunological synapses (10).
Alternate Names
KPCT; MGC126514; MGC141919; nPKC-theta; PKC theta; PKC-theta; PKCQ; PKCT; PKCtheta; PRKCQ; PRKCT; protein kinase C theta; Protein kinase C theta type; protein kinase C, theta
