Recombinant full-length human CAMK2α was expressed by baculovirus in Sf9 cells using an N-terminal GST tag. CAMK2α is a ser/thr protein kinase that is a member of the Ca2+/calmodulin-dependent protein kinase family. CAMK2α is abundant in the brain as a major constituent of the postsynaptic density and is required for hippocampal long-term potentiation (LTP) and spatial learning. In addition to its Ca2+/calmodulin-dependent activity, CAMK2α can undergo autophosphorylation, resulting in Ca2+/calmodulin-independent activity. The protein level of CAMK2α fluctuates during neuronal activity in cultured rat pup hippocampal neurons. The levels of CAMK2α increased with heightened neuronal activity (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: CAMK2α, 10μg (Human, recombinant full-length). MW: ~74kDa. Substrate: Autocamtide 2 peptide (KKALRRQETVDAL-amide); derived from the autophosphorylation site (amino acids 283–290) on CaMKII. Other: Reaction Buffer, DTT, Ca2+/Calmodulin solution.
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.Silva, A.J. et al. (1992) Impaired spatial learning in alpha-calcium calmodulin kinase II mutant mice. Science 257, 206–11.
2.Thiagarajan, T.C. et al. (2002) Alpha- and beta-CaMKII: inverse regulation by neuronal activity and opposing effects on synaptic strength. Neuron 36, 1103–14.