Structure of His-Tag complex
Darren Tyson
tysondr at slu.edu
Thu May 8 12:04:20 EST 1997
Thorsten Schmidt wrote:
>
> Dear reader!
>
> The His-Tag-System is widely used for protein purification.
> (Novagene, Quiagen ...)
>
> Using a special expression vector, a "His-Tag", a repeat of six Histidines,
>
> is added to the target protein.
> A special resin is then loaded with nickel-ions.
> When one loads the column with a protein mixture only
> the target protein forms with its histidines a complex with the nickel and
> the resin.
> After washing steps the protein can be eluted with a buffer containing
> Imidazole.
>
> I want to know the exact structure of this complex and the reason, why
> it is possible to elute the protein with Imidazole.
>
> Can you help me, perhaps tell me a reference?
>
> Thank you very much in advance for your answer
>
> Thorsten
Thorsten,
QIAGEN's web site has plenty of information available regarding
purification of His-tagged proteins. In particular, they have an online
version of the QIAexpressionist which is a great resource for the theory
behind the expression and purification of recombinant His-tagged
proteins. The URL is
http://www.qiagen.com/qexpress/startqxp.html
Hope this helps,
Darren
**********
Darren Tyson
Ph.D. Candidate in Cell & Molecular Biology
Saint Louis University
tysondr@<spam_blocker>slu.edu
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