Product Name: PKCq-pY545
Product Number: AB-PK773
Size: 25 µg      Price:89.00
      $US
Target Full Name: Protein-serine kinase C theta

Target Alias: Kinase PKC-theta; KPCT; MGC126514; MGC141919; NPKC-theta; PKCQ; PKC-theta; PRKCQ; PRKCT; Protein kinase C, theta; Protein kinase C, theta type; CCDS7079.1; Q04759; ENSG00000065675

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK773

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y545

Protein UniProt: Q04759

Protein SigNET: Q04759

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human PKCt (PRKCQ) sequence peptide Cat. No.: PE-04ADG99

Antibody Immunogen Sequence: TPD(pY)IAP(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues T542 to P548; In the protein kinase catalytic domain activation T loop region between subdomains VII and VIII.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 0.75 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 75-84 kDa.

Related Product 1: PKCq-pY545 blocking peptide

Related Product 2: PKCq-pS695 phosphosite-specific antibody (Cat. No.: AB-PK772)

Scientific Background: PKCt (PKCq, PRKCT, PKC-theta) is a protein-serine/threonine kinase of the AGC group and novel PKC family. It is dependent on acidic phospholipids (e.g. phosphatidylserine) and diacylglycerol for full phosphotransferase activity, and does not require calcium. It is essential in TCR signalling for T-cell activation, differentiation, proliferation, and survival. These T-cell responses are mediated via Jun, NF-kappa-B, NFATC1 and NFATC2. The phosphotransferase activity of PKCt is unaffected by an Y90F mutation, but T-cell proliferation is inhibited. An A148E mutation results in constitutive activation of PKCt. A T219A mutation again will not inhibit kinase phosphotransferase activity, but it will inhibit localization to the plasma membrane and inhibit transactivation of IL2 promoter. Phosphorylation at Y90, T538 (activation loop of the kinase domain) and S676 (turn motif) increases phosphotransferase activity. Phosphorylation at S695 (hydrophobic region) increases phosphotransferase activity and interaction with PDK1. Insertional mutagenesis studies in mice support a role for this protein kinase in mouse cancer oncogenesis. PKCt is linked to Gastrointestinal Stromal Tumours (GANT, GIST), which are rare sarcomas arising from tissues lining the gastrointestinal tract and typically arise from the stomach, intestines, or colon. GANT has been related to neurofibromatosis and adenocarcinoma as well.
Figure 1. Dot blotting PKCq-pY545 antibody with recombinant purified proteins.


Figure 2. Epitope mapping of PKCq-pY545 antibody with similar phosphopeptides on dot blots.


Figure 3. Identification of phosphosites related to PKCq-pY545.