ADP-Glo Kinase Assay + MST1 Kinase Enzyme System
Recombinant full-length human MST1 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. MST1 belongs to a family of proteins that share similarity with a budding yeast serine/threonine kinase, sterile-20 (Ste20). Endogenous full-length MST1 is activated by a variety of stressful stimuli, accompanied by the secondary appearance of a 36kDa Thr183-phosphorylated, caspase-cleaved catalytic fragment (1). Recombinant MST1 undergoes a robust auto-activation in vitro, mediated by an intramolecular autophosphorylation on the activation loop of an MST dimer. MST1 can initiate apoptosis when transiently overexpressed in mammalian cells. Interference with the ability of endogenous MST1 to associate with the putative tumor suppressor proteins Nore1/RASSF can inhibit Ras-induced apoptosis (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: MST1, 10ug (Human, recombinant full-length). MW: ~83kDa. Substrate: Axltide (KKSRGDYMTMQIG); derived from mouse Insulin receptor substrate 1 (amino acids 979-989). Other: Reaction Buffer, DTT. MST1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/6789/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.