Recombinant full-length human AMPK (combination of A1/B1/G2 subunits) was expressed by baculovirus in Sf9 insect cells using a C-terminal His tag. AMP-activated protein kinase (AMPK) exhibits a key role as a master regulator of cellular energy homeostasis (1). AMPK exists as a heterotrimeric complex composed of a catalytic alpha subunit and regulatory beta and gamma subunits. Binding of AMP to the gamma subunit allosterically activates the complex. AMPK is activated in response to stresses that deplete cellular ATP (low glucose, hypoxia and ischemia) (2) and via signaling pathways in response to adiponectin, leptin and CAMKKbeta. 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: AMPK (A1/B1/G2), 10ug (Human, recombinant full-length). MW: ~68kDa (A1); ~38kDa (B1); ~65kDa (G2). Substrate: SAMStide (HMRSAMSGLHLVKRR); derived from the mouse acetyl-Coenzyme A carboxylase alpha (amino acids 73-85). Other: Reaction Buffer, DTT, AMP Solution. AMPK (A1/B1/G2) NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/5562/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.|
Profile More Compounds In-House: ADP-Glo(TM); 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(TM); Kinase Assay result in less ambiguous data.