Description
Background/Introduction: Phosphatidic Acid (PA) is a critical precursor for the biosynthesis of many lipids in the cell. PA plays a critical role in membrane structure and acts as a signaling lipid, recruiting cytosolic proteins to the cell membrane. Within the cell, PA concentrations are maintained at extremely low levels by the activity of potent phospholipid phosphatases, converting phosphatidic acid to diacylglycerol (DAG). Because DAG is another important lipid precursor, it too is rapidly metabolised into other membrane lipid components (e.g. phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine). Ultimately, measurement of cellular PA is useful for monitoring lipid synthesis and metabolism. Total Phosphatidic Acid Assay Kit measures total phosphatidic acid content, including lysophosphatidic acid (LPA), in samples by a coupled enzymatic reaction system. First, lipase is used to hydrolyze phosphatidic acid in samples to glycerol-3-phosphate. Next, 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 Total Phosphatidic Acid Assay Kit is a simple, fluorometric assay that quantitatively measures total PA (PA and LPA) in samples using a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 100 assays, including blanks, standards and unknown samples. The kit contains an L-alpha-Phosphatidic Acid Standard and has a detection sensitivity limit of ~5 uM