ADP-Glo Kinase Assay + DAPK1 Kinase Enzyme System
Recombinant human DAPK1 (1-363) 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 gamma-interferon. Activation of DAPK occurs via dephosphorylation of Ser-308 and subsequent association of calcium/calmodulin (1). 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 (2). 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 chemical compounds have 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. Kinase Enzyme System contains: Kinase: DAPK1, 10ug (Human, recombinant; amino acids 1-363). MW: ~71kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT, Ca2+/Calmodulin Solution. DAPK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/1612/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.