native HisTag elution alternative

R. John Lye rjl6n at virginia.edu
Fri Oct 27 07:04:25 EST 2000


soenke behrends wrote:

> Dear netters,
>
> we purify a protein containing a HisTAG with
> a Ni-NTA matrix and elute the protein with
> imidazole (250mM). Unfortunately enzyme activity
> of the eluted protein is strongly inhibited by
> 250mM or 150mM imidazole, whereas 50mM imidazole
> lead to 50% of the activity without imidazole.
>
> We now want to turn to an alternative elution
> approach

Is there a reason that you can't dialyze the imidazole away?

John







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