Recombinant full-length human DGKA was expressed by baculovirus in Sf9 insect cells using an N-terminal His tag. Diacylglycerol kinase alpha (DAG kinase alpha) is a member of the eukaryotic diacylglycerol kinase family. Upon cell stimulation it converts the second messenger diacylglycerol into phosphatidate, initiating the resynthesis of phosphatidylinositols and attenuating protein kinase C activity. Involved in hepatocellular carcinoma progression through regulation of the Ras/Raf/MEK/ERK pathway. 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.
