SYK Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The ZAP70 Kinase Enzyme System contains:

• ZAP70 Kinase, 10μg (Human, recombinant full-length). MW: ~96kDa.
• Poly (4:1 Glu, Tyr) Peptide Substrate.
• Reaction Buffer, DTT, MnCl₂.

Recombinant full-length human ZAP70 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ZAP70 is a nonreceptor protein tyrosine kinase (part of the Syk/Zap70 family) that is involved in signaling by the T-cell antigen receptor (TCR). Ligation of the TCR/CD3 receptor in Jurkat T-cells induces phosphoprotein complexes, which contain ZAP70. TCR zeta chains are initially phosphorylated by p56Lck that lead to the recruitment of ZAP70 via its SH2 domain. ZAP70 in turn phosphorylates other proteins in the TCR-phosphoprotein complex. One of the natural substrates for ZAP70 is the zeta-chain dimer of the TCR/CD3 complex.

ZAP70 NCBI Database Entry.

The ZAP70 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.