T cell specific kinase

Accession #/Synonyms: BAA02873, IL2-inducible T cell kinase, Kinase EMT, T cell specific  kinase,  TSK.

Protein kinases constitute a large superfamily of enzymes with key regulatory functions in nearly all signal transmission processes of eukaryotic cells. Signal transduction through the T-cell receptor and cytokine receptors on the surface of T lymphocytes occurs largely via tyrosine phosphorylation of intracellular substrates. The Tec family Tyrosine kinase ITK has been implicated in T cell receptor (TCR) signaling. The tec family kinase, inducible T cell tyrosine kinase (Itk), is critical for both development and activation of T lymphocytes. The TCR-CD3-induced actin-dependent cytoskeletal events are regulated by ITK phosphorylation.  The ITK kinase activity is increased upon its phosphorylation by Lck at tyrosine 511 (1). 

The peripheral T cell lymphomas (PTCL) unidentified (PTCLU) represent the largest PTCL. Most patients with advance PTCLU present with frequent involvement of extranodal sites including spleen, bone marrow, liver, blood.  These patients also exhibit a chromosomal translocation t(5;9)(q33;q22) which fuses  the ITK and spleen tyrosine kinase (SYK) genes, both these kinases are required for normal antigen-induced lymphocyte activation (2). The generation of constitutively active fusion tyrosine kinases (FTKs)2 by chromosomal translocation is a recurrent mechanism of oncogenesis in hematological neoplasms.  The ITK possess an amino terminal plecktrin homology (PH) domain for plasma membrane recruiting, a Tec homology domain, Src homology (SH) 3 and SH2 domains for association with T cell adapter proteins and kinase domain for TCR downstream signaling (3). The chromosome translocation in PTCLU fuses the PH and TH regions form ITK to the SYK kinase domain that causes aggressive T cell lymphomas (3).  The ITK is a 620 aminoacid (75kda) protein, the ITK gene resides on chromosome 5q33.   

ITK-selective antibodies were generated against peptides form unique region near C-terminal end of ITK protein, corresponding to amino acid 573-594. This region was not only antigenic but also lies on the surface of the protein in native or in partially denatured confirmation.  The ITK antibodies are fully characterized for applications listed here at the recommended antibody dilutions.  The ITK antibodies are affinity purified on an immobilized antigen based affinity matrix, the isolated antibodies were then stabilized in antibody stabilization buffer for long-term storage.  FabGennix Int. Inc. provides ITK Western blot positive control samples in “ready-to-use” SDS-PAGE sample buffer.  Limited quantities of antigenic blocking peptide for ITK is also available, please inquire before ordering.  FabGennix Int. Inc., will also conjugate this antibody to fluorophores and other secondary enzymes at a nominal price. 

Immunogen:  Synthetic peptide taken from human ITK protein taken from within the C-terminal domain of ITK corresponding to aa: tgfrlykprlasthvyqimnhc. The peptide was conserved in several other species.  The selected peptide was post-synthetically modified to achieve highest antigenicity before coupling to carrier protein using hetero bifunctional cross linker for immunogen preparation.                 

Concentration: Affinity purified TBK1-101AP antibody (0.54-0.57mg/ml) was stabilized in antibody stabilization buffer for long-term storage.

Applications:   ITK-101AP antibody was tested in ELISA and western blotting applications at 1:500 dilution using PC-Slc1A samples.  Antibody dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions.  Application of this antibody in other protocols has not yet tested. WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d.  Investigators using this antibody in protocols other than listed above can request a complimentary sample of this antibody.

Protocols:       Standard protocol for various applications (WB, IMM, IHC) of this antibody can be obtained upon request. The specification sheet for TBK1 antibody will be supplied with each product.  FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Reactivity:  The antibody labels a strong band of Slc1A 78kDa in PC-ITK sample. The antibody also labels a weak band of 69kDa.  The identity of this band is not known but could represent a proteolytic product of ITK..  

For research use only, not for diagnostic or therapeutic use.