His-tag binding resins
hgtsei at med-rz.uni-sb.de
Tue Jun 10 08:19:57 EST 1997
In article <339C600B.3B43 at post.queensu.ca>, carstens at post.queensu.ca wrote:
> Sorry if this has been posted before but we are just starting out with
> some His-tagged constructs and want to purify the expressed proteins.
> Are there any self-made columns and purification protocols available
> anywhere that we can try out, rather than just buying the stuff from
> Thanks for any help!
I would recommend Talon-Agarose sold by Clontech. It uses Co2+ rather
than Ni2+-ions and shows somewhat less co-purified proteins upon
purification. However, in my hands pure recombinant proteins in one step
were only obtained from bacterial cultures and with the aid of a chromato-
graphic pump system capable of producing gradients (of imidazole).
If you express proteins from eucaryotic cells, it is almost impossible
to get pure proteins with just one affinity step. Here, Co2+-Agarose
is superior to Ni2+.
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