To go along with the EDTA comment, did you check the pH of your binding solution? Sometimes lysing E. coli causes the pH to drop below the pKa of the histidines.
On Tue, Jan 27, 2009 at 8:10 PM, James Stroud <xtald...@gmail.com> wrote: > You don't have EDTA left over from your protease inhibitor, do you? Some > commercial cocktails include it. You may have to read the fine print. > > James > > > On Jan 27, 2009, at 1:00 PM, Fred wrote: > > Hi ccp4 list, >> I am trying to purify a his-tag protein by metal affinity chromatography. >> The protein was expressed in inclusion bodies and its his-tag doesn't bind >> the Qiagen Ni resin in denatured conditions (urea 8M or GndHCl 6M). Playing >> with NaCl and detergents didn't help much. >> Any help is appreciated. >> Fred >> > -- Jim Fairman Graduate Research Assistant Department of Biochemistry, Cellular, and Molecular Biology (BCMB) University of Tennessee -- Knoxville 216-368-3337 jfair...@utk.edu james.fair...@case.edu
