refolding on IMAC-column

Antonin Tutter atutter at aim.salk.edu
Mon Jul 27 22:59:21 EST 1998


>Hi netters,
>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?
>Christian


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

_______________________________________
Antonin Tutter
Salk Institute for Biological Studies
RBIO-J
10010 N. Torrey Pines Rd.
La Jolla, CA  92037
email:  atutter at aim.salk.edu
web:  http://www-biology.ucsd.edu/~atutter/




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