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.
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.Okuno, S. et al. (2001) Regulation of Ca2+/calmodulin-dependent protein kinase kinase alpha by cAMP-dependent protein kinase: I. Biochemical analysis. J. Biochem (Tokyo) 130, 503–13.
2.Hurley, R.L. et al. (2005) The Ca2+/calmodulin-dependent protein kinase kinases are AMP-activated protein kinase kinases. J Biol Chem. 280, 29060–6.