Re.:How to renature His-tagged protein

Roger Murphy roger.murphy at ludwig.edu.au
Wed May 9 01:45:42 EST 2001


Try running a decreasing gradient of urea 8 to 0M over say 5-10 column volumes
before you elute your protein from the Ni-NTA column.  I hear this results in
pretty good folding but haven't tried it personally...

Bull Zebra wrote:

> I am purifying a 6XHis-tagged protein under denature condition (8M urea)
> using Ni-NTA agarose and I need to renature the protein in order to couple
> it to an affinity column (CNBr method). Anybody there can suggest a good
> renaturation protocol for this purpose? Thanks in advance!
>
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