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The enzyme reactions contained 50 μl of the diluted supernatant, 450 μl of 0.04 mol/L phosphate buffer, and 3.6 ml substrate (α-naphtyl acetate), which were mixed and incubated in a shaker for 30 min at 37°C

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Plus, older adults are more likely to have pernicious anemia or atrophic gastritis, which also affect B12 levels