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The redox behavior of the NAD + /NADH system is therefore influenced by 1) the pH of the solution, since hydrogen ions participate in the redox half-reaction, 2) the 8 protonation states of NAD + , due to its three basic moieties, 3) the 16 protonation states of NADH, due to its four basic moieties (two pyrophosphate moieties, an adenine ring, and an added dihydropyridine ring)
In both cases, gently swirl the bottle to homogenizenever shake vigorously to avoid denaturation or oxidation