IUBio

6x his tag protein purification

Tom Belbin tbelbin at morgan.ucs.mun.ca
Fri Mar 26 11:53:43 EST 1999


In article <7dg5fs$utt$1 at news.iastate.edu>, "Dept of Microbiology"
<bfeilmei at iastate.edu> wrote:

> Help!!
> 
> I have cloned GFP behind a 6X his tag to purify the protein.  I am binding
> it to TALON Cobolt beads.  The protein almost binds too well, in that, I am
> having a lot of trouble eluting it off.  I have heard rumor of someone using
> histidine in the elution buffer to compete for the removal of the his tagged
> protein.  Does anyone know of a good way to elute hard to remove his tagged
> proteins?
> 
> I appreciate the help!!

Many protocols use imidazole to compete with the tag for binding to the column.
Have you tried imidazole?

Tom Belbin
Biochemistry Department
Memorial University of Newfoundland




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