NADP+/NADPH Colorimetric Assay Kit (WST-8) (E-BC-K803-M)
For research use only.
| Detection Principle |
Detect total content of NADP+ and NADPH: Glucose 6-phosphate (G6P) is oxidized to 6-phosphate gluconolactone (6-PG) by glucose-6-phosphate dehydrogenase (G6PDH), and NADP+ is reduced to NADPH during this reaction. NADPH, under the action of 1-mPMS, transfer electrons to WST-8 to produce the yellow product, which has a characteristic absorption peak at 450 nm. Therefore, the total content of NADP+ and NADPH can be quantified by measure the OD value at 450 nm. Detect NADPH: After treating sample, heat at 60℃ water bath for 30 min. the NADP+ of the sample is decomposed and only NADPH remains. NADPH reduces WST-8 to form Formazan, and the amount of NADPH is determined by measure the OD value at 450 nm. Detect NADP+ and NADP+/NADPH: The content of NADP+ and the ratio of NADP+/NADPH in the sample can be obtained according to the total content of NADP+ and NADPH obtained of the first two steps as well as the separate content of NADPH. |
| Synonyms | NADP;NADPH |
| 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 , Cuproptosis , Disulfidptosis |
| Other Reagents Required | Ultrapure water |
| 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: 5.5% | intra-assay CV: 2.1% |
| Sample Volume | 50-200 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 |
|---|---|
| Jurkat cells | 1 |
| Mark cells | 1 |
| HCT116 cells | 1 |
| 293T cells | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| Hela cells | 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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