I was told by a lab colleague that 'after purifying a protein with a his-tag using imidazole, it is crucial to quickly remove the imidazole. Keeping the protein in more than 200mM of imidazole is a very bad idea, and storing the protein in over 200mM of imidazole can also affect its activity.'
So, what is the reason that imidazole should be removed quickly, and what is the mechanism by which imidazole causes the protein to lose its activity?
Adam B Shapiro
As far as I know, it is not necessary to quickly remove the imidazole, as long as the pH of the elution buffer is suitable for the stability of the protein. Be sure to adjust the pH of the imidazole-containing buffer before using it to elute the protein.
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