NAD(P)H-Glo™ Detection System Technical Manual
Literature # TM398
The NAD(P)H-Glo™ Detection System is a homogeneous bioluminescent assay that generates a light signal from biochemical reactions that contain reduced nicotinamide adenine dinucleotide (NADH) or nicotinamide adenine dinucleotide phosphate (NADPH). The NAD(P)H-Glo™ Detection System quantitatively monitors the concentration of the reduced forms, NADH and NADPH, and does not discriminate between them. The oxidized forms, NAD+ and NADP+, are not detected and do not interfere with quantitation. The assay can be used to monitor the activity of enzymes that produce or use NADH and NADPH and to screen for inhibitors. In the presence of NADH or NADPH, the enzyme Reductase reduces a proluciferin reductase substrate to form luciferin. Luciferin then is quantified using Ultra-Glo™ Recombinant Luciferase, and the light signal produced is proportional to the amount of NADH and NADPH in the sample. The assay is compatible with 96-, 384-, low-volume 384- and 1536-well plates and is well suited to measure NADH and NADPH production or consumption in high-throughput formats.
Summary of Changes
The following changes were made to the 8/17 revision of this document:
1. The recommended storage temperature of unused Reductase Substrate (Section 3.B, Step 5) was changed.
2. Related Products (Section 7) was updated to include new products.