protein folding on Ni-column

D.K. at
Thu Sep 4 15:01:09 EST 2003

In article <20030904190629.17359.qmail at>, richardfundgrueb at 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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