NADPH Fluorometric Assay Kit (E-BC-F170)
For research use only.
| Detection Principle | Glucose-6-phosphate (G6P) is oxidized by glucose-6-phosphate dehydrogenase (G6PDH) to gluconolactone-6-phosphate (6-PG). At the same time, NADP+ is reduced to NADPH in this reaction. The generated NADPH reacts with a chromogenic agent under enzymatic catalysis to produce a fluorescent substance. The fluorescence intensity can be measured to calculate the NADPH content in the sample. |
| Sample Type | Animal tissue,Cell |
| Detection Method | Fluorometric method |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Other Reagents Required | Double distilled water, PBS (0.01 M, pH 7.4) |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid Period | 12 months |
| Sensitivity | 8.55 nmol/L |
| Detection Range | 8.55-1000 nmol/L |
| Precision | inter-assay CV: 3.3%-8.4% | intra-assay CV: 1.7%-5.0% |
| Sample Volume | 100-200 μL(tissue/cell homogenate) |
| Assay Time | 2 h |
The recommended dilution factor for different samples is as follows (for reference only):
| Sample Type | Dilution Factor |
|---|---|
| 10% Mouse liver tissue homogenization | 1 |
| 10% Mouse heart tissue homogenization | 1 |
| 1×10^6 RAW264.7 cells | 1 |
| 1×10^6 HL 60 cells | 1 |
| 1×10^6 A549 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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