Determination of Significance:
Coenzyme II NADP(H) is widely present in animals, plants, microorganisms and cultured cells. The determination of NADP+ and NADPH content can calculate the content of NADP (NADPH + NADP+) and the ratio of NADPH/NADP+, and its changes are related to the pentose phosphate pathway and biosynthesis and Antioxidative responses are closely related. The NADPH/NADP+ ratio is not only one of the main markers of cellular redox state, but also plays an important regulatory role in the PPP pathway, biosynthesis and antioxidant metabolism.
Measurement Principle:
The NADP+ and NADPH in the samples were extracted with acidic and basic extraction solutions, respectively. Under the action of 1-mPMS, WST-1 can react with NADPH to produce water-soluble formazan, which has a characteristic absorption peak at 450nm, while NADP+ can be reduced to NADPH by 6-phosphate glucose dehydrogenase, which is further detected by WST-1.
Self Provided:
Spectrophotometer, centrifuge, water bath, mortar/homogenizer, sonicator, adjustable pipette, 1 mL glass cuvette, ice, and distilled water.
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