Description:
Pyridine nucleotides play an important role in metabolism and, thus, there is continual interest in monitoring their concentration levels. Quantitative determination of NAD+/NADH has applications in research pertaining to energy transformation and redox state of cells or tissue. Simple, direct and automation-ready procedures for measuring NAD+/NADH concentration are very desirable. This NAD+/NADH assay kit is based on a lactate dehydrogenase cycling reaction, in which the formed NADH reduces a formazan (MTT) reagent. The intensity of the reduced product color, measured at 565 nm, is proportional to the NAD+/NADH concentration in the sample. This assay is highly specific for NAD+/NADH and with minimal interference (<1%) by NADP+/NADPH. Our assay is a convenient Method to measure NAD, NADH and their ratio.