PEP is a key intermediate in glycolysis and gluconeogenesis and an essential component for glucose homeostasis. Cayman's Phosphoenolpyruvate Assay provides a fluorescence-based method for measuring PEP in red blood cells and tissue homogenates. In this assay, pyruvate kinase catalyzes the transfer of a phosphate group from PEP to ADP, yielding one molecule of pyruvate and one molecule of ATP. Pyruvate oxidase then catalyzes the conversion of pyruvate to acetyl phosphate, hydrogen peroxide (H2O2), and carbon dioxide. H2O2, in the presence of horseradish peroxidase, reacts stoichiometrically with ADHP (10-acetyl-3,7,-dihydroxyphenoxazine) to produce the highly fluorescent compound resorufin, which can be detected with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.