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I am having a similar problem All Answers (11) If the protein is on the column but won't elute even with 50 mM GSH, chances are it has precipitated on the column

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In addition to biological activity as an essential intracellular antioxidant, GSH facilitates the clearance of metabolic waste via GSH- S -transferases and may function as a neuropeptide

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When 1ml makes sense The 1ml reconstitution works best in specific situations