His-tagged proteins....

Michael Witty mw132 at mole.bio.cam.ac.uk
Fri Mar 23 05:29:04 EST 2001

. . . a lot of people would say, don't bother removing the tag if you can
demonstrate activity with the tag present.  Can you easily do that?  Mike.

On 22 Mar 2001, Debashis Mukhopadhyay wrote:

> Hi All!
> This question is directed to those 'gurus' who have
> expertise with His-tagged proteins to help me out of
> this problem. I'm new to this "affinity tagged"
> protein field.
> I have a protein domain which is cloned and expressed
> in E.coli having an N-terminal His-tag and a Thrombin
> cleavage site just after that. The idea is to get rid
> of the tag for crystallisation purposes by thrombin
> treatment. Can you point me to any reference(s), ideal
> for a novice, having details for the same and
> subsequent purification steps to eliminate the
> contaminants! Any nice protocol would as well be good!
> Any help would be highly appreciated.
> Regards,
> Debashis
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