DAG Kinase Activity (DAG) Assay Kit

Background/Introduction: Diacylglycerol (DAG) and phosphatidic acid (PA) are key molecules in the biosynthesis of many cellular lipids and play fundamental roles in biochemical signaling. Diacylglycerol acts as a precursor to many lipids (e.g. triglycerides, phospholipids); however, DAG also functions as a second messenger signaling lipid, produced through hydrolysis of PIP2 by phospholipase C, which initiates intracellular Ca2+ release and PKC activation. Diacylglycerol kinase (DAGK) is a family of enzymes that phosphorylate diacylglycerol to yield phosphatidic acid, utilizing ATP as a source of phosphate. Therefore, DAGK is thought to be a key regulating enzyme of numerous cellular responses by modulating the balance between these two signaling lipids. 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, a lipase is used to hydrolyze phosphatidic acid to glycerol-3-phosphate. Finally, the glycerol-3-phosphate product is oxidized by glycerol-3-phosphate oxidase (GPO), producing hydrogen peroxide which reacts with the kit's Fluorometric Probe (Ex. 530-560 nm/Em. 585-595 nm). The DAG Kinase Activity Assay Kit is a non-isotopic, fluorometric assay that quantitatively measures DAGK activity using its physiological substrate (diacylglycerol) in a 96-well microtiter plate format. Recombinant DAGK is also provided as a positive control. Each kit provides sufficient reagents to perform up to 50 assays, including blanks, standards and unknown samples.

Small volumes of DAG assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS169290 is an assay kit targeting DAG. Also known as DAG Kinase Activity Assay Kit.