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Procedure Preheat spectrophotometer/microplate reader for 30 minutes, adjust the wavelength to 412 nm, set zero with distilled Preheat Reagent II in 37 (mammal cell) or 25 (other species) water bath for 30 Blank tube determination: take micro glass cuvette, add 20 L of distilled water, 140 L of Reagent II, 40 L of Reagent III in turn, mix well, place for 2 minutes, and measure 412 nm absorbance AB