refolding on IMAC-column
atutter at aim.salk.edu
Mon Jul 27 22:59:21 EST 1998
>i want to refold a recombinant produced protein on an IMAC column by
>decreasing step-by-step the urea concentration. In some papers I have
>read that it´s important to avoid high protein concentartions. But this
>is the case when I bind my protein to the gel matrix. Have anyone of you
>experience with refolding on a IMAC column?
I have successfully refolded proteins from an IMAC column by first eluting
at low pH (Qiagen has a nice protocol for this). This reduces the problem
of localized concentration peaks on the column. I then refolded by
sequencial dialysis (pH 7.9, 1M urea steps down to 1M, then 0.5M, then 0M).
Hope this helps
Salk Institute for Biological Studies
10010 N. Torrey Pines Rd.
La Jolla, CA 92037
email: atutter at aim.salk.edu
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