ADP-Glo Kinase Assay + CAMKK1 Kinase Enzyme System
Recombinant full-length human CAMKK1 was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. CAMKK1 or CAMKK alpha is a Ca2+/calmodulin-dependent protein kinase that activates CAM-kinases I and IV via phosphorylation of their Thr(177) and Thr(196) residues, respectively. Recent studies have shown that the activity of CAMKK1 is decreased upon phosphorylation by cAMP-dependent protein kinase (PKA) (1). CAMKKI has been identified in intact cells as AMPKKs, predicting a significant role for this kinase in regulating AMPK activity in vivo. It has been shown that 2-deoxyglucose- and ionomycin-stimulated AMPK activity is substantially reduced in HeLa cells transfected with small interfering RNAs specific for CAMKKI (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: CAMKK1, 10ug (Human, recombinant full-length). MW: ~94kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT, Ca2+/Calmodulin Solution. CAMKK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/84254/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.