Purification with His-Tag

Sigrid Van Boxstael svboxsta at vub.ac.be
Tue Mar 28 02:48:00 EST 2000


Frank Fuerst wrote:

> Anton Tutter schrieb:
> >
> > >> Can I remove the Imidazol just by dialyzing?
> > >
> > > Yes, we usually do that.
> >
> > just a side warning about dialyzing the imidazole away from his-tag
> > proteins--his tagged proteins have a tendency to aggregate and precipitate
> > out of solution during the dialysis step.  this has been an ongoing topic of
> > discussion on this newsgroup.
>
> And what would you recommend instead? Are there other possibilities to
> get rid of imidazole, where the aggregation problem is minimized?

I dialyzed my protein and indeed it  precipitated in the dialyses bag.
Maybey it is possible to get rid of the imidazol by doing a gelfiltration.
Or do you think the proteine might precipitate on the column?

Sigrid

Sigrid Van Boxstael
VUB MICROBIOLOGIE
E. Gryzonlaan 1
1070 Brussels
Belgium







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