protein folding on Ni-column
no.email at thanks.to.spam.net
Thu Sep 4 15:01:09 EST 2003
In article <20030904190629.17359.qmail at ww02.hostica.com>, richardfundgrueb at hotmail.com wrote:
>I want to refold a protein bound by His-tag to a Ni-column. Can anyone give me
> some references in which this has been previously done?
No refs that I have on hand but depending on the protein, one of the
two extreme approaches should work:
1. Quickly dump and mix your sepharose-bound protein into
100-300 column volumes of a solution without denaturant and let
sit for an hour or so.
2. Run slow (20-30 column volumes) gradient from 100% to 0%
denaturant through a column.
Both done better at RT but there are, I believe, some examples
of #2 working better in the cold room.
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