Roger Murphy roger.murphy at
Fri Feb 23 00:10:22 EST 2001

Tertiary amines such as Tris can strip metal ions from the column - change
your buffers.


vladimir vigdorovich wrote:

> Hi, I am trying to optimize a purification procedure of a His-tagged
> protein derived from Hi5 cells using Talon resin. When I apply clarified,
> pH-adjusted (to ~8.0), buffered (supplemented with 20 mM Tris-HCl) supe to
> the column, it slowly changes color to whitish-pink and the flowthrough
> darkens. Some protein binds but I suspect that much ends up in
> flowthrough.
> Can anyone verify my suspicions that Co2+ is being leeched and/or suggest
> a remedy?
> Thanks,
> Vlad
> vladv at

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