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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

Apostolopoulos V, McKenzie IF (1994) Cellular mucins: targets for immunotherapy

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