Total Phosphatidic Acid Assay Kit (Fluorometric)User Manual
|Research Area:||Lipid Metabolism|
|Assay Category:||Phospholipid Assays|
|Sample Type:||Detects total phosphatidic acid content (PA and LPA) in cell lysate samples|
Phosphatidic Acid (PA) is a phospholipid that serves three main roles in the cell: it is a precursor for lipid biosynthesis, provides membrane structure, and acts as a signaling lipid by recruiting cytosolic proteins to the cell membrane. PA concentrations are maintained at low levels in the cell by phospholipid phosphatases, which convert PA to diacylglycerol (DAG). Measurement of cellular PA levels is useful for monitoring lipid synthesis and metabolism. Our Total Phosphatidic Acid Assay Kit measures total phosphatidic acid content (PA and LPA) in cell lysate samples by a coupled enzymatic reaction system. First, lipase is used to hydrolyze phosphatidic acid to glycerol-3-phosphate. Next, the glycerol-3-phosphate product is oxidized by glycerol-3-phosphate oxidase (GPO), 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. Phosphatidic Acid levels in unknown sampled are determined based on the provided phosphatidic acid standard curve.