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+. The changes are related to the pentose phosphate pathway and biosynthesis and resistance The oxidation reaction is closely related. The ratio of NADPH/NADP+ is not only one of the main signs of cell redox state, but also has an important regulatory role in PPP pathway, biosynthesis and antioxidant metabolism.
Measurement Principle:
The NADP+ and NADPH in the sample were extracted with acidic and alkaline extracts, respectively. NADPH reduces the oxidized thiazole blue (MTT) to formazan through the hydrogen transfer effect of PMS, and detects the absorbance at 570nm to determine the NADPH content. Use 6-phosphate glucose dehydrogenase to reduce NADP+ to NADPH to detect NADP+ content.
Self Provided:
Spectrophotometer/microplate reader, centrifuge, water bath, mortar/homogenizer, sonicator, adjustable pipette, micro glass cuvettes/96 well plates , ice, and distilled water.
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