Dialyse

Randy rldurren at bchserver.bch.ncsu.edu
Fri Nov 3 11:09:31 EST 2000


I found that DTT will turn the His-bind-nickel column matrix orange (VERY
difficult to remove). We dialyze out of the imidazole, with very good
results.
Randy

"Martin Offterdinger" <martin.offterdinger at akh-wien.ac.at> wrote in message
news:8tu713$599o$1 at www.univie.ac.at...
>
> "Frank Fuerst" <ffrank at rz.uni-potsdam.de> schrieb im Newsbeitrag
> news:8ts15v$22ue$1 at fu-berlin.de...
> > "Richard P. Grant" <rpg14 at yahoo.co.uk.invalid> wrote:
> >
> > >In article <8tq8il$8nq$1 at news.doit.wisc.edu>,
> > >klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) wrote:
> > >
> > >> The nickel is gone. We are talking about _recharging_ and that
> > >> necessarily implies stripping the original, fully or partially
reduced,
> > >
> > >From Cornelius' posting, that's not that crystal clear.   I'm not sure
> > >whether he reduced it in the nickel-bound state or not.
> >
> > I would even say that it _is_ clear that the nickel was still there.
> > The original poster wanted an alternative for imidazole as an eluent,
> > and asked wether EDTA or DTT would work:
> >
> > >We now want to turn to an alternative elution
> > >approach e.g. EDTA. [...]
> > >
> > >I have also been thinking about whether one could
> > >use high concentrations of DTT. In the manual
> > >(Quiaexpressionist) they write that DTT can reduce
> > >Nickel thereby preventing nickel from binding His-tagged
> > >proteins.
> >
> > And Cornelius warned him to use DTT.
> >
> > Bye, Frank
>
> What about dialysing out the imidazole?
> martin
>
>







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