| Intrument | Microplate reader (450 nm) |
| Assay Time | 70 min |
| Validity | 6 |
| Assay Type | Quantitative |
| Sample Type | animal tissu, cells |
| Synonyms | NADP/NADPH |
| Detection Principal | 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. |
| Reagents | Ultrapure water |
| Labware | Test tube, Micropipettor, 37? water bath, 10 KD filters tube. |
| Size | 96T |
| Sensitivity | 0.02 ?mol/L |
| Detection Range | 0.02-5.0 ?mol/L |
| Inter CV | 5.5 |