Recombinant full-length human DGKZ was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Diacylglycerol kinase zeta (DAG kinase zeta) is a member of class IV diacylglycerol kinase family, which contains two zinc fingers, an ATP binding site, and four ankyrin repeats with the MARCKS phosphorylation site, and exhibits the preference for 1,2-diacylglycerols but not for long chain diacylglycerols. DGK zeta regulates Ras activation via terminating the diacylglycerol signaling induced RasGRP. 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.
