For assessing PDH activity in biological samples and screening and evaluation of PDH modulators.

• High sensitivity and wide linear range
• The detection limit is 0.87 U/L, with linearity up to at least 278 U/L PDH in a 96-well plate assay
• Homogeneous and simple procedure
• Simple 'mix-and-measure' procedure allows reliable quantitation of PDH activity within 20 minutes
• Robust and amenable to HTS
• All reagents are compatible with high-throughput liquid handling instruments

PYRUVATE DEHYDROGENASE (PDH) (E.C. 1.2.4.1) is an oxidoreductase which catalyzes the decarboxylation of pyruvate to form acetylated dihydrolipoamide in a thiamine pyrophosphate (TPP)-dependent reaction. PDH is the E1 component of the PDH complex that ultimately produces acetyl-CoA. The PDH complex connects glycolysis to the citric acid cycle. This non-radioactive colorimetric PDH assay is based on the reduction of the tetrazolium salt MTT in a Phenazine Methosulfate (PMS) coupled reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The intensity of the purple color formed is directly proportional to the PDH activity.