Phospholipase D hydrolyzes phosphatidylcholine to yield choline and phosphatidic acid (PA), which plays a role in intracellular signaling, mitogenesis in fibroblasts, stimulation of respiratory bursts in neutrophils, increase of cellular Ca2+, and activation of specific protein kinases and phospholipases. Cayman's Phospholipase D Assay provides a convenient fluorescence-based method for determining PLD activity in both crude and purified enzyme preparations. The assay detects choline molecules which are generated during the hydrolysis of phosphatidylcholine. Choline is oxidized by Choline Oxidase to produce betaine and hydrogen peroxide (H2O2). In the presence of peroxidase, H2O2 reacts with 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) to yield the highly fluorescent compound resorufin. Resorufin fluorescence can be analyzed using an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.