NAD/NADH-Glo™ Assay Technical Manual
Literature # TM399
The NAD/NADH-Glo™ Assay is a bioluminescent homogeneous assay for detecting total oxidized and reduced nicotinamide adenine dinucleotides (NAD+ and NADH, respectively) and determining their ratio in biological samples. In the presence of NAD+ and NADH, 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 NAD+ and NADH in the sample. The assay has a linear range of 10nM to 400nM NAD+ and high signal-to-background ratios and is specific for the nonphosphorylated forms. The assay is compatible with 96-, 384-, low-volume 384- and 1536-well plates and is well suited to monitor the effects of small molecule compounds on NAD+ and NADH levels in enzymatic reactions or directly in cells in high-throughput formats.
Summary of Change
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 6) was changed.
2. Related Products (Section 9) was updated to include new products.