Phospholipase D Assay

Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine to generate phosphatidic acid, and subsequently releasing soluble choline. PLD plays an important role in a variety of cellular functions and has been associated with the pathogenesis of diseases such as cancer, Alzheimer's disease, Parkinson's disease, and brain disorders. In this colorimetric assay, phosphatidylcholine is converted to choline by PLD, where choline is then oxidized by choline oxidase and the formed hydrogen peroxide is catalyzed by peroxidase. This product reacts with 4-aminoantipyrine to form a colored solution that can be read at 550nm. The intensity of the colored reaction product is directly proportional to the activity of PLD in the sample.