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protein purification/ endotoxinremoval
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Posted 23 March 2001 - 10:00 PM
I am trying to purify a His-tagged protein that is secreted into cell culture medium (AIM V) by a human cell line. The problem is that the cell culture medium seems to strip the Nickel from the resin. Is thereany way to circumvent this problem other then dialysis, which considering the volume of medium (about 2 litres) seems rather impractical. I don't want to precipitate the protein, since this would necessitate refolding. Would addition of nickel to the culture medium help? If anyone has any ideas I would bevery grateful. Thanks.