How to renature His-tagged protein

jose jose at biochemistry.cwru.edu
Tue May 8 17:30:26 EST 2001

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!

many years ago we used to purify RNAP components using denaturation, and
then applying them to an ion exchange column (DEAE) and then, as they
elute in conditions that strongly favor folding, they would refold as
they returned to the bulk solution.

maybe a variation on that theme as one approach?

jose nazario				jose at cwru.edu

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