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.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.De Souza, P.M. et al. (2004) Mammalian Sterile20-like kinase 1 and the regulation of apoptosis. Biochem Soc Trans. 32, 485–8.
2.Avruch, J. et al. (2005) Nore1 and RASSF1 regulation of cell proliferation and of the MST1/2 kinases. Methods Enzymol. 407, 290–310.