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Pyruvate Kinase Activity Assay Kit

A fluorescence-based method for detecting PK activity in erythrocyte lysates, plasma, cell culture samples, and tissue homogenates

Pyruvate kinase (PK) catalyzes the formation of pyruvate and ATP from phosphoenolpyruvate and ADP. Deficient expression of PK in erythrocytes is one common cause of hemolytic anemia, and because it is expressed in tumor cells, PK has been studied as a potential tumor target. Cayman's Pyruvate Kinase Activity Assay provides a fluorescence-based method for detecting PK activity in erythrocyte lysates, plasma, cell culture samples, and tissue homogenates. In this assay, PK 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 10-acetyl-3,7,-dihydroxyphenoxazine (ADHP) 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.