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Pyruvate Dehydrogenase (PDH) Assay Kit

Components:

PDH Assay Solution: 5 ml; store at -20°C (for 100 assays)

10X Cell Lysis Solution: 25 ml; store at 4°C

Product Description: Pyruvate is the end product of aerobic glycolysis and is at an important metabolic crossroad. The pyruvate dehydrogenase multi-enzyme complex (PDH) catalyzes the oxidative decarboxylation of pyruvate to acetyl-CoA in a series of reactions mediated by the E1 subunit (pyruvate dehydrogenase), E2 subunit (dihydrolipoyl transacetylase), and E3 subunit (dihydrolipoyl dehydrogenase). The reactions of the PDH complex serve to interconnect the metabolic pathways of glycolysis, gluconeogenesis and fatty acid synthesis to the Krebs (TCA) cycle. As a consequence, the PDH complex is highly regulated by a variety of allosteric effectors and by covalent modification. The importance of the PDH complex in metabolic regulation is evident from the fact that although diseases associated with deficiencies of the PDH complex have been observed, affected individuals often do not survive to maturity. This non-radioactive colorimetric PDH assay is based on the reduction of the tetrazolium salt INT in a NADH-coupled enzymatic reaction to formazan, which is water-soluble and exhibits an absorption maximum at 492 nm. The intensity of the red color formed is increased in the presence of increased PDH activity. The kit is stable for at least one year under proper storage and handling conditions. The product is for research use only.

Features: The non-radioactive assay kit allows the assay to be completed within 30-60 min with minimal pipetting steps. Each kit is sufficient for approximately 100 assays using the 96-well format. See related product LDH assay kit, GAPDH assay kit, G6PD assay kit, Glucose assay kit, and Lactate Assay Kit. The method is adaptable to automation.