It should work however the question of protein stability in these
conditions is still there.
I would try EDTA first, unless you know that your protein contains
structurally vital metal ions.
Also, why can you try Ni-NTA ? It has higher affinity for Ni ions and
might not be leached out by histidine.
A.
Jeremy Murray wrote:
>> What about low pH or EDTA?
>> John Eisses wrote:
>> > 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? I
> > seem to remember a ref for N acetyl Histidine , but does this strip the
> > Talon resin? I'll try buffer exchange after the fact but I'd like to not
> > lose activity as my experience is once its gone it doesn't come back.
> >
> > salud
> > Ted Michelini
> > michelini at ohsu.edu
--
|Dr. Artem Evdokimov Protein Engineering |
| NCI-Frederick Tel. (301)846-5401 |
| FAX (301)846-7148 |
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