Diacylglycerol (DAG) is a glyceride consisting of two fatty acid chains linked by a central glycerol backbone. It is a precursor to lipids such as triglycerides and phospholipids and functions as a second messenger signaling lipid. It is produced through hydrolysis of PIP2 by phospholipase C, which initiates intracellular Ca2+ release and PKC activation. Activated PKC enzymes interact with many different substrates, affecting cellular physiology.
Our DAG (Diacylglycerol) Assay Kit measures diacylglycerol in cell lysate samples by a coupled enzymatic reaction system. First, kinase is used to phosphorylate DAG in samples, 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. The hydrogen peroxide released from this reaction reacts specifically with the kit’s Fluorometric Probe and is detected at ex. 530-560 nm/em. 585-595 nm. DAG levels in unknown samples are determined based on the provided DAG standard curve.