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Furthermore, hemolysis as an ACD is non-specific and reflects only the mechanistic damage to cells
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The supernatant was collected and analyzed for NADP + and NADPH in three steps as follows: (a) a spectrophotometric reading was taken using untreated extract to determine total amount of NADPH and NADH in the sample (absorbance value A1 at 340 nm), (b) an aliquot of the extract was incubated with G6PDH (Sigma) to convert all of the NADP + in the sample to NADPH, and its levels were determined as in step 1 to produce absorbance value A2, (c) another aliquot of the extract was incubated with glutathione reductase (Calbiochem) to convert all of NADPH to NADP + , and its levels were determined as in Steps 1 and 2 to obtain absorbance value A3
