Product Name:
ITK-pY512
Product Number:
ab-pk666
Target Full Name: Tyrosine-protein kinase ITK/TSK
Target Alias: EMT; Lyk; PSCTK2; MGC126257; MGC126258; TLK; TSK; CCDS4336.1; ENSG00000113263
Product Type Specific: ITK protein kinase phosphosite-specific antibody
Antibody Code: PK666
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y512
Protein UniProt: Q08881
Protein SigNET: ITK
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. Y512 phosphorylation stimulates phosphotransferase activity. This phosphosite is located in the kinase activation loop between catalytic subdomains VII and VIII. This is the major in vivo phosphorylation site in ITK (≥256 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). ITK is known to be phosphorylated at this site in vitro by Lck.
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.
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: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Mouse | Platypus | Chicken | Zebra fish | Fruit fly.
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 (509-515) pY512 antibody blocking peptide (Cat. No.: PE-04ANB90)
Related Product 2: ITK-BCT pan-specific antibody (Cat. No.: AB-NK274-1)
Scientific Background: ITK is a 72 kDa non-receptor tyrosine kinase that is a member of the TK group of protein kinases in the Tec family. It features SH2, SH3, Pleckstrin Homology (PH), and Tec homology (TH) domains, which are crucial for membrane recruitment and protein interactions. It is primarily especially expressed in T lymphocytes, essential for T-cell receptor (TCR) signaling, development,and differentiation. ITK is essential for Th2 differentiation, cytokine production (IL-4, IL-5, IL-13), and actin cytoskeletal rearrangement. It acts as a mediator for immune responses, with deficiencies linked to allergies, infections, and malignancies. While primarily in T-cells, ITK is also found in Natural Killer (NK) cells and mast cells. It 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 Tyr-12. ITK require prior activation of Lck, ZAP70 and PI3-kinase for efficient activation. Activated ITK phosphorylates Phospholipase C-gamma1 (PLCG1), triggering calcium release and activating the nuclear factor of activated T-cells (NFAT) for gene transcription. 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). This description may include information annotated by UniProt and/or Google AI.