Diacylglycerol (DAG) and phosphatidic acid (PA) are key molecules in the biosynthesis of many cellular lipids and play fundamental roles in biochemical signaling. DAG Kinase (DAGK) phosphorylates DAG to yield PA, using ATP as a source of phosphate. DAGK is thought to be a key regulating enzyme of numerous cellular responses by modulating the balance between these two signaling lipids.
Our DAG Kinase Activity Assay Kit measures DAGK activity in samples by a coupled enzymatic reaction system. First, kinase-containing sample is used to phosphorylate DAG substrate, yielding phosphatidic acid. Next, lipase is used to hydrolyze phosphatidic acid to glycerol-3-phosphate. The glycerol-3-phosphate product is then oxidized by glycerol-3-phosphate oxidase, producing hydrogen peroxide which reacts specifically with the kit’s Fluorometric Probe.