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Protein was eluted with 4CV of 60% buffer B and immediately diluted 10-fold with Buffer A prior to concentration and 3 rounds of buffer exchange to remove excess imidazole by centrifugation at 3095xg at 4 C in 100 kDa cutoff concentrators (Amicon)

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B12 injections circumvent these pathways entirely, delivering the full dose directly into the bloodstream where it is absorbed with near-complete efficiency

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The antioxidant function of GSH is determined by the redox-active thiol (-SH) of cysteine that becomes oxidized when GSH reduces target molecules (Pompella et al., 2003)