Product Name: ITK-pY512
Product Number: AB-PK666
Size: 25 µg      Price:89.00
      $US
Target Full Name: Tyrosine-protein kinase ITK/TSK

Target Alias: EMT; Lyk; PSCTK2; MGC126257; MGC126258; TLK; TSK; CCDS4336.1; ENSG00000113263

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK666

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y512

Protein UniProt: Q08881

Protein SigNET: Q08881

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human ITK sequence peptide Cat. No.: PE-04ANB90
Antibody Immunogen Sequence: DDQ(pY)TSS(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues D509 to S515; In protein kinase catalytic domain activation T-loop 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: 1 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 Potency: Weak immunoreactivity of a target-sized protein by Western blotting in Jurkat cells. Very strong immunoreactivity with immunogen peptide on dot blots.

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 68-72 kDa.

Antibody Specificity: High

Antibody Cross Reactivity: No significant cross-reactivities detected in Jurkat cells, except phenylarsine oxide (PAO) increases 15 kDa protein immunoreactivity.

Related Product 1: ITK-pY512 blocking peptide

Related Product 2: ITK-BCT pan-specific antibody (Cat. No.: AB-NK274-1)

Scientific Background: ITK is a protein-serine/threonine kinase that is a member of the TK group of protein kinases in the Tec family. This kinase is widely distributed from moderate to high levels in most human tissues with highest expression in bladder, lung, lymph node, spleen, stomach, thymus, tonsils and uterus. Ligation of CD2, TCR or CD28 induces activation and tyrosine phosphorylation of ITK at Y512. ITK is expressed in T-cells and is important for T-cell development and activation through the antigen receptor. ITK require prior activation of Lck, ZAP70 and PI3-kinase for efficient activation. ITK knockout mice show multiple effects on T cell development, cytokine production and T-helper cell differentiation. T cells that lack or express mutant versions of ITK show impaired TCR-induced actin polymerization, cell polarization and regulation of the signalling events involved in cytoskeletal reorganization. ITK has been linked with the development of gastric adenocarcinomas, melanoma (metastatic) and lymphoproliferative syndrome ebv-associated autosomal type 1 (LPSA1).
Figure 1. Epitope mapping of ITK-pY512 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to ITK-pY512.