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8 mM of reducing agent DTT (Sigma) was then added to the samples and incubated in a thermomixer at 30C for 30 minutes, followed by the addition of 20 mM iodoacetamide (Sigma) for protein carboxylation with incubation at 30C in the dark for 30 minutes

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The higher the initial pool and the more efficient the person is at recycling and conserving cobalamin, the longer it will take for the deficiency to develop