ANTI-ITK CLONE 2F12

General description

Tyrosine-protein kinase ITK/TSK (EC 2.7.10.2; UniProt Q08881; also known as IL-2-inducible T-cell kinase, Interleukin-2-inducible T-cell kinase, Kinase EMT) (T-cell-specific kinase, Tyrosine-protein kinase Lyk) is encoded by the ITK (also known as EMT, LPFS1, LYK) gene (Gene ID 3702) in human. Interleukin-2-inducible T-cell kinase (Itk) is a TEC family non-receptor tyrosine kinase that plays an important roles in signal transduction in T-cells. Upon cell surface receptor stimulation, Itk is recruited to the membrane by the adaptor molecule LAT (linker for activated T cells), where Itk is transphosphorylated at Tyr511 by Lck, Itk Tyr180 then takes place for full activation. Once activated, Itk transduces cellular signaling by activating phospholipase Cγ1 (PLCγ1), leading to activation of nuclear factor of activated T-cells (NFAT), nuclear factor κB (NFκB), and mitogen-activated protein kinase (MAPK) pathways. Itk-/- mice are resistant to developing ovalbumin-induced asthma and exhibit reduced cytokines production by iNKT cells, as well as reduced Th2 effector cytokines and IL-17A productin by T-cells.

Specificity

Clone 2F12 targets the N-terminal region of human, mouse, and rat Itk.

Immunogen

GST-tagged recombinant human/mouse/rat Itk N-terminal fragment.

Application

Detect ITK using this mouse monoclonal Anti-Itk Antibody, clone 2F12, Cat. No. MABS1281, validated for use in Immunoprecipitation and Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A 1:1000 dilution from a representative lot detected Itk in 10 µg of HuT-78 human T-cell lymphoma cell lysate.

Immunoprecipitation Analysis: A representative lot immunoprecipitated Itk from both parental and Lck-negative (J.CaM1) Jurkat cells. Itk immunoprecipitated from the parental Jurkat cells was found with increased tyrosine phosphorylation following TCR cross-linking or pervanadate treatment, while only very weak tyrosine phosphorylation was seen with Itk from J.CaM1 upon prolonged pervanadate treatment (Heyeck, S.D., et al. (1997). J. Biol. Chem. 272(40):25401-25408).

Immunoprecipitation Analysis: A representative lot immunoprecipitated tyrosine-phosphorylated Itk from Jurkat cells stimulated by cross-linking of endogenous cell surface CD3 or transfected CD8 extracellular domain/CD28 cytoplasmic domain chimera (Heyeck, S.D., et al. (1997). J. Biol. Chem. 272(40):25401-25408).

Western Blotting Analysis: A representative lot detected baculovirus expressed Itk from infected Sf9 insect cells following ltk immunoprecipitation by the same antibody. The overexpressed ltk was found autophosphorylated on tyrosine when expressed alone in Sf9 and increased ltk tyrosine phosphorylation was seen when co-expressed with Lck (Heyeck, S.D., et al. (1997). J. Biol. Chem. 272(40):25401-25408).

Quality

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Itk in 10 µg of Jurkat cell lysate.

Target description

~71 kDa observed. 71.83/72.29/72.49 (human/mouse/rat) kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.