Recombinant full-length human DGKB was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Diacylglycerol kinase beta (DAG kinase beta) is a member of class I diacylglycerol kinase family and is regulated by Ca2+ via converting diacylglycerol into phosphatidic acid, diacylglycerol kinases decrease the diacylglycerol levels to regulate the activity of PKC, Rho and Ras families. DGK beta is localized in brain and is involved in emotion and long-term memory linked to cognitive function. 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.
