Pyruvate Kinase Activity Assay

Pyruvate kinase (PK) is a key enzyme in the glycolytic pathway catalyzing the conversion of phosphoenolpyruvate to pyruvate forming ATP. This reaction serves as a key control step regulating the overall rate of glycolysis and determining the fate of glucose metabolism. Pyruvate kinase plays a central role providing energy for cellular processes, as pyruvate is a metabolite common to several pathways. Pyruvate assay helps in understanding glycolytic flux, assessing enzyme regulation, and evaluating metabolic alterations in diseases including cancer, diabetes, or genetic disorders. AkrivisBio’s Pyruvate Kinase Activity Assay is a simple, sensitive assay for determining PK activity in a variety of sample types below 50 μU. In the assay, pyruvate kinase converts Phosphoenolpyruvate and ADP to generate pyruvate and ATP. Pyruvate is then oxidized by pyruvate oxidase, forming hydrogen peroxide. Hydrogen peroxide is utilized by peroxidase to oxidize ADHP resulting in the formation of intense color/fluorescence measured at OD 570 nm or at Ex/Em = 535/587 nm respectively.