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Invitrogen™ NAD+/NADH Colorimetric Assay Kit, 96 Tests
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Specifications:
| Application | Colorimetric Assay | ||
| Storage Temperature | 2-8°C | ||
| Product Type | Assay Kits | Forms | Kit with Various components |
| Product Brand | Thermo Fisher Scientific™, Invitrogen | ||
| Product Grade | Molecular Biology | ||
The Invitrogen™ NAD⁺/NADH Colorimetric Assay Kit is a sensitive and reliable solution for the quantitative determination of total NAD⁺ and NADH, individual NADH levels, and calculation of the NAD⁺/NADH ratio in biological samples. This kit is designed to support research in cellular metabolism, mitochondrial function, and redox biology.
The assay is based on an enzyme-coupled reaction in which NADH reduces WST-8 to form a stable yellow formazan product with a characteristic absorbance at 450 nm, allowing accurate quantification using a standard microplate reader.
Importantly, NADP⁺ and NADPH do not interfere with the assay, ensuring specificity for NAD⁺/NADH measurements.
Assay Principle
1. Total NAD⁺ + NADH Measurement
Ethanol is enzymatically converted to acetaldehyde, during which NAD⁺ is reduced to NADH. The generated NADH transfers electrons to WST-8, producing a yellow-colored product measured at 450 nm.
2. NADH Measurement
Samples are heated at 60 °C to selectively decompose NAD⁺, leaving NADH intact. NADH then reduces WST-8, allowing direct determination of NADH content.
3. NAD⁺ and NAD⁺/NADH Ratio
By subtracting NADH from total NAD⁺ + NADH, the NAD⁺ concentration and NAD⁺/NADH ratio can be calculated.
Key Features
- Simple & Fast Workflow – Minimal hands-on time with rapid incubation
- High Sensitivity – Detects NAD⁺/NADH down to 0.02 μmol/L
- Specific Detection – No interference from NADP⁺/NADPH
- Quantitative & Reproducible – Low intra- and inter-assay variability
- Broad Application Scope – Suitable for metabolic and redox studies
- Consistent Performance – Manufactured under strict quality control
Applications
- Cellular metabolism research
- Redox biology and oxidative stress studies
- Mitochondrial function analysis
- Energy metabolism and aging research
- Pharmacological and toxicological studies
Technical Specifications
| Parameter | Specification |
|---|---|
| Assay Range | 0.02 – 5.0 μmol/L |
| Assay Sensitivity | 0.02 μmol/L |
| Detection Method | Colorimetric |
| Detection Wavelength | 450 nm |
| Total Assay Time | ~70 minutes |
| Incubation Time | 30 minutes |
| Sample Volume | 20 μL |
| Sample Type | Animal tissue, cells |
| Target Species | Universal |
| Equipment Required | Microtiter plate reader |
| Intra-assay CV | 1.8% |
| Inter-assay CV | 9.1% |
| Kit Format | Colorimetric detection kit |
| Unit Size | Each (96 tests) |
Contents & Storage
- Kit Contents: Optimized reagents for NAD⁺/NADH detection (see kit insert)
- Storage Conditions: 2 °C to 8 °C
- Shipping Conditions: Dry ice, 2 °C to 8 °C
The Invitrogen™ NAD⁺/NADH Colorimetric Assay Kit (EEA042) offers a robust, sensitive, and user-friendly platform for studying cellular energy metabolism and redox balance. With excellent specificity, reproducibility, and compatibility with standard laboratory equipment, it is an ideal choice for researchers investigating NAD⁺ dynamics, metabolic regulation, and mitochondrial health.