Phoebe, We were able to purify the estrogen receptor DNA binding domain with a 6his-tag. After cutting off the tag and re-applying to the Ni-NTA matrix, the protein did not stick to the beads. We had some low resolution crystals that contained a second protein and DNA, which suggests to me that the zinc was bound in the protein, but we did not analyze it further.
Kendall -- Kendall W. Nettles, PhD Assistant Professor Department of Cancer Biology The Scripps Research Institute 5353 Parkside Dr. Jupiter Fl 33458 office 561-799-8851 fax 561-799-8805 cell 561-306-7566 On 1/3/08 7:21 PM, "[EMAIL PROTECTED]" <[EMAIL PROTECTED]> wrote: > This has probably been discussed before, so apologies in advance. > We're eyeing a protein that has a probable C4 Zn finger in the > middle. The collaborators who are nicely going to PCR it up want to > know if we'd like it with or without a His tag. > Is it a bad idea to co-mingle Zn-binders and Ni columns? Or is it > likely to bind the column quite nicely without the tag? > thanks, > Phoebe > > > ------------------------------------------------------------------------------ > --------------------------------------------- > Phoebe A. Rice > Assoc. Prof., Dept. of Biochemistry & Molecular Biology > The University of Chicago > phone 773 834 1723 > fax 773 702 0439 > http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabe > tically.php?faculty_id=123 > http://www.nasa.gov/mission_pages/cassini/multimedia/pia06064.html
