Determination of NADH Concentrations with the SynergyTM 2 Multi-Detection Microplate Reader using Fluorescence or Absorbance

Many biological assays have as their basis a link to the oxidative status of nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP). Many dehydrogenase enzymes use these coenzymes to transfer hydrogen groups between molecules. Because the reduced forms of these molecules differ from the oxidized forms in their ability to absorb light, it is possible to quantitate reactions based on light absorbance at 340 nm or by the fluorescent emission of light at 445 nm. Here we describe the use of the SynergyTM 2 Multi-Detection Microplate Reader to quantitate NADH using either fluorescence or absorbance modes.