NAD+/NADH Colorimetric Assay Kit (WST-8) (E-BC-K804-M)
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For research use only.
| Detection Principle |
Detect total content of NAD+ and NADH: Ethanol generates acetaldehyde under the action of enzyme. Meanwhile, NAD+ is reduced to NADH, NADH, under the action of hydrogen transmitter, transfer electrons to WST-8 to produce the yellow product, which has a characteristic absorption peak at 450 nm. Therefore, the total content of NAD+ and NADH can be quantified by measure the OD value at 450 nm. Detect NADH: After treating sample, heat at 60oC water bath for 30 min. the NAD+ of the sample is decomposed and only NADH remains. NADH reduces WST-8 to form yellow product, and the amount of NADH is determined by measure the OD value at 450 nm. Detect NAD+ and NAD+/NADH: The content of NAD+ and the ratio of NAD+/NADH in the sample can be obtained according to the total content of NAD+ and NADH obtained of the first two steps as well as the separate content of NADPH. Note: NADP+ and NADPH have no effect on the determination results. |
| Synonyms | NAD+/NADH |
| Sample Type | Animal tissue,Cell |
| Detection Method | Colorimetric method |
| Detection Instrument | Microplate reader (450 nm) |
| Research Area | Oxidative Stress , Metabolic Diseases , Glycolysis And Lipid Metabolism , Energy Metabolism |
| Other Reagents Required | PBS (0.01 mol/L, pH 7.4) |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid Period | 12 months |
| Sensitivity | 0.02 μmol/L |
| Detection Range | 0.02-5.0 μmol/L |
| Precision | inter-assay CV: 9.1% | intra-assay CV: 1.8% |
| Sample Volume | 20 μL(tissue/cell homogenate) |
| Assay Time | 1 h 10 min |
The recommended dilution factor for different samples is as follows (for reference only):
| Sample Type | Dilution Factor |
|---|---|
| 10% Mouse muscle tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 293T cell | 1 |
| Hela cell | 1 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
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