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Assuming you followed the above advice and used an appropriate binding buffer and your protein isnt huge, then a likely reason for this is that the expression yield is much lower than you expected

Both the number and size of adenomas were significantly reduced in patients given a high dose of aged garlic extract compared to those given a much lower dose (0.16 mL/day) (131, 132)

Elements that influence the treatments success include: Liver Health: Since the liver produces glutathione naturally, a healthier liver supports better absorption and utilization

While hGH did produce a slight increase in glycerol in the knockout mice, the response was markedly lower than in WT animals