SV: SV: Talon resin histidine use
Dag Rune Gjellesvik
dagrg at online.no
Sat May 5 07:55:02 EST 2001
I've used up to 0.1 M glycine on zink-loaded chelate sepharose (APBiotech)
without any leakage. I think that glycine has a lot lower chelating activity
than imidazole, but all this depends on which side binds the metal best -
the matrix side, or the his-tag on the protein side.
Michael Witty <mw132 at mole.bio.cam.ac.uk> skrev i
meldingsnyheter:Pine.SGI.4.33.0105031556520.26400027-100000 at mole.bio.cam.ac.
> . . . but wouldn't that leach cobalt from the column matrix? Mike.
> On Thu, 3 May 2001, Dag Rune Gjellesvik wrote:
> > Hi,
> > I'm not sure of the chemistry in the Talon resin, but you may try 0.1 -
> > M glycine. Glycine has mild chelating activity and may release the
> > without releasing the cobalt.
> > Dag Rune
> > John Eisses <eissesj at ohsu.edu> skrev i
> > meldingsnyheter:3AEEF610.5D9FEE48 at ohsu.edu...
> > > Bionetters,
> > > I am purifying a delicate membrane protein and imidazole seems to
> > > inhibit it, but when I use histidine, the Talon (Co2+) resin bleaches
> > > out and the buffer turns brown. Are there alternatives for imidazole?
> > > seem to remember a ref for N acetyl Histidine , but does this strip
> > > Talon resin? I'll try buffer exchange after the fact but I'd like to
> > > lose activity as my experience is once its gone it doesn't come back.
> > >
> > > salud
> > > Ted Michelini
> > > michelini at ohsu.edu
> > >
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