The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The ITK Kinase Enzyme System contains:
ITK Kinase, 10μg (Human, recombinant; amino acids 352–end). MW: ~53kDa.
Poly (4:1 Glu, Tyr) Peptide Substrate.
Reaction Buffer, DTT, MnCl2.
Recombinant human ITK (amino acids 352–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ITK is a member of the TEC family of nonreceptor tyrosine kinases. ITK is expressed in T-cells and is important for T-cell development and activation through the antigen receptor. ITK requires prior activation of Lck, Zap-70 and PI3-kinase for efficient activation and shares major substrates with both Lck and Zap-70. 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 signaling events involved in cytoskeletal reorganization.
ITK NCBI Database Entry.
The ITK Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.
Use with ADP-Glo™ Kinase Assay
The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.
See all Kinase Enzyme Systems available from Promega.