His6-Tagged Protein: Binding strength, panning...
jb261 at cryst.bioc.cam.ac.uk
Wed Mar 4 12:20:48 EST 1998
I've pulled interacting proteins out before using a 6-His tagged protein on
Qiagne NTA-Ni beads. To prevent non-specific interactions I used 20 mM
imidazole and 300 mM KCl in the buffer. I found that I got the best
results when I ran the column continuously (i.e. recycling flow through)
over night and then eluting the next day.
S. Findley wrote:
> Ok I have a his6-tagged protein which I would like to immobilize on a
> Ni-NTA column and subsequently bind and purify interacting proteins by
> passing a cell extract over the column.
> Is this a generally used approach?
> Are there preferable tags to immobilize proteins on columns?
> (Is th affinity of His6 for the Ni sufficient for this type of protocol?
> Seth D. Findley polliwog at u.washington.edu
> Department of Biochemistry Phone: 206-543-1710
> Health Sciences Building J579
> University of Washington, Box 357350, Seattle, WA 98195
Department of Biochemistry
Cambridge CB2 1QW
ph. 01223 766027
More information about the Methods