Description
The in vitro enzyme-based NAD(P)H-Glo™ Detection System is a homogeneous, bioluminescent assay that quantitatively monitors the concentration of the reduced forms of nicotinamide adenine dinucleotides, NADH and NADPH, and does not discriminate between them. The oxidized forms, NAD+ and NADP+, are not detected and do not interfere with quantitation.
In the presence of NAD(P)H, a reductase enzyme 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(P)H in the sample. The reductase and luciferase reactions are initiated by adding an equal volume of a single reagent, which contains reductase, proluciferin Reductase Substrate and Ultra-Glo™ Recombinant Luciferase, to a NAD(P)H-containing sample.
The assay is rapid, requiring only a 40- to 60-minute incubation, has a broad linear range and high signal-to background ratio. The assay is well suited to measuring NAD(P)H production or consumption in high-throughput formats.
Features – Benefits
- Broad Linear Range: The NAD(P)H-Glo™ Detection System detects 0.1μM to 25μM NAD(P)H.
- High Sensitivity: The limit of detection is ≤0.1μM NADH, with a maximum assay window (i.e., signal-to-background ratio) of 250. The system detects 1μM with a signal higher than fivefold over background.
- Automation Compatible: The add-and-read format is compatible with automated and high-throughput protocols. Reactions are scalable and can be performed at low volumes in 96-, 384- and 1536-well plates.
- Reliability and Reproducibility: The NAD(P)H-Glo™ Detection System routinely yields Z´ factors >0.7.
- Stable Signal: The glow-type signal is stable, with a half-life greater than two hours, allowing batch plate processing.
- Luminescence-Based NAD(P)H Detection: The luminescent format avoids fluorescent interference due to reagents and test compounds sometimes seen in fluorescent assays.
Applications
- Monitoring the activity of enzymes that produce or use NAD(P)H.
- Measuring NAD(P)H production or consumption in high-throughput formats.
| Product | Size | Brand | Catalog |
| NAD(P)H-Glo™ Detection System | 10 ml | Promega | G9061 |
| NAD(P)H-Glo™ Detection System | 50 ml | Promega | G9062 |
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