Pyruvate dehydrogenase (PDH) is the first component enzyme of the pyruvate dehydrogenase complex (PDC). It catalyzes the conversion of pyruvate to acetyl-CoA and CO₂, thereby linking the tricarboxylic acid (TCA) and glycolysis pathways. Mutations in PDH have been linked to pyruvate dehydrogenase deficiency (causing lactic acidosis and neurologic dysfunctions) and Leigh syndrome. PDH has also been implicated in oncogene-induced senescence. Measuring PDH activity can provide insights into metabolic functions and oncogenesis.

Pyruvate Dehydrogenase Activity Assay Kit provides a simple, reproducible, and sensitive tool for measuring pyruvate dehydrogenase activity in cell lysates, plasma, serum, urine, and other biological liquid samples. This assay features low disturbance and high sensitivity. It 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.

• Microplate reader capable of measuring absorbance at 566 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Multi-channel pipettes to deliver 20 µL to 200 µL volumes
• Tubes to prepare sample dilutions
• 50 mL conical tubes
• Ultrapure water

1. Prepare Working Solution.
2. Prepare Positive Control.
3. Add 20 µL of the diluted PDH Positive Control, Assay Buffer (used as a blank) or unknown samples to a 96-well microtiter plate.
4. Add 100 µL of Working Solution to each well. Mix thoroughly. Read and record absorbance at 566 nm every minute for 3 time points at room temperature (25°C).