ADP-Glo Kinase Assay + TOPK 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 DAPK1 Kinase Enzyme System contains:

• DAPK1 Kinase, 10μg (Human, recombinant; amino acids 1363). MW: ~71kDa.
• Native Swine Myelin Basic Protein (MBP) Substrate.
• Reaction Buffer, DTT, Ca²⁺/Calmodulin Solution.

Recombinant human DAPK1 (1363) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Death-associated protein kinase 1 (DAPK1) is a positive mediator of apoptosis induced by γ-interferon. Activation of DAPK occurs via dephosphorylation of Ser-308 and subsequent association of calcium/calmodulin. DAPK is rapidly dephosphorylated in response to tumor necrosis factor or ceramide and then subsequently degraded via proteasome activity. The decline in DAPK expression is paralleled with increased caspase activity and cell apoptosis. Studies suggest that the apoptosis regulatory activities mediated by DAPK are controlled both by phosphorylation status and protein stability.

DAPK1 NCBI Database Entry.

The DAPK1 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.