NAD/NADH Assay Kit

Pyridine nucleotides play an important role in metabolism, so there is interest in monitoring their concentration levels in metabolic systems. Quantitative determination of NAD+/NADH has applications in research pertaining to energy transformation and the redox state of cells or tissue. Nicotinamide adenine dinucleotide (NAD) is an enzymatic cofactor that plays a pivotal role in numerous redox reactions. Serving as an electron carrier, NAD seamlessly transitions between its oxidized form (NAD) and reduced form (NADH). Apart from its involvement in redox reactions, NAD assumes essential roles in ADP-ribosylation reactions and serves as a substrate for sirtuins.

The kit is suitable for NAD+/NADH concentration and ratio determination in cell or tissue extracts.

The 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 with minimal interference (<1%) by NADP+/NADPH. The assay is a convenient method to measure NAD, NADH, and their ratio.