Itk (IL-2 Inducible T-cell Kinase, Emt, Tsk, T Cell Signaling Protein)

Itk is a T cell signaling protein. The Tec family of non-receptor tyrosine kinases is composed of six proteins designated Tec, Itk (also known as Emt or Tsk), Btk (previously known as Atk, BPK or Emb), Bmx, Txk (also known as Rlk) and Dsrc28C. All members of the family contain SH3 and SH2 domains and, with the exception of Txk and Dsrc28C, also contain pleckstrin homology (PH) and Tec homology (TH) domains in their amino termini. Tik associates with CD28 and becomes activated subsequent to CD28 ligation. \n\nApplications:\nSuitable for use in Western Blot, Immunoprecipitation and Protein Kinase Assay. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 1:1000-1:2000 dilution detects Itk in RIPA lysates from Jurkat cells. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with I9200. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.\nImmunoprecipitation: 4ul immunoprecipitated Itk from 500ug of Jurkat RIPA lysate.\nProtein Kinase Assay: Reported to immunoprecipitate active Itk.4.\n\nPositive Antigen Control Included: \nJ9000: Jurkat cell lysate. Add 2.5ul of 2- mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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