Imidazol induced oligomerization?

Christian Radauer rc150295 at emb1.bcc.univie.ac.at
Fri Dec 11 13:38:06 EST 1998


Dima Klenchin wrote:
> 
> In article <366D1A38.9FF502B9 at pasteur.fr>, Pierre Rodrigues <pirod at pasteur.fr> wrote:
> >Hello,
> >
> >Does anybody knows if imidazol (used as eluant of his tag purification)
> >can induce or at least strongly stabilize protein oligomerization?
> >
> >This because when I purify a receptor by histag technique I always
> >obtain it as if it was an oligomer (detected by western-blot), even
> >resistant to SDS-PAGE. When I look at it directly by western-blot on
> >crude extracts I have it mainly as a monomer with some higher bands. So,
> >could imidazol have stabilize the interaction?
> 
> NO.

Hi!

Indeed it is not the imidazol that causes oligomerization, but the
His-tag, which is crosslinked by heavy metal ions. I have not tried it
yet, but maybee adding EDTA to the PAGE sample buffer would help.

Christian Radauer
Institute of General and Experimental Pathology
University of Vienna, Austria



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