The NAD(P)H-Glo™ Detection System is a bioluminescent, homogeneous, single-reagent-addition assay that quantitatively monitors the concentration of the reduced forms of NADH and NADPH levels in enzymatic reactions. The oxidized forms, NAD+ and NADP+, are not detected and do not interfere with quantitation.
The assay is rapid, requiring only a 40- to 60-minute incubation, has a broad linear range (25nM-50µM) and high signal-to background ratio (~400). Low nanomolar sensitivity, 25nM, allows direct in-well detection. The assay is well suited to measuring NAD(P)H production or consumption in high-throughput formats.
In the presence of NAD(P)H, a reductase reduces the proluciferin reductase substrate to form luciferin. Luciferin is quantified using Ultra-Glo™ Recombinant Luciferase, which is a component of the Luciferin Detection Reagent, and the light signal produced is proportional to the amount of NAD(P)H in the sample.