His-tagged proteins

Julie jroberts at rascal.med.harvard.edu
Fri May 9 10:21:56 EST 1997


I'm currently trying to work through this myself. Upon calling Novagen
tech support the gave those suggestions but also suggested using a
decreasing pH gradient to elute the protein. At least with the system
I'm using the pH is near 8 throughout the procedure.

Julie Roberts
jroberts at rascal.med.harvard.edu



Cara Baker wrote:
> 
> Has anyone had experience with his-tagged proteins that precipitate
> immediately on elution with imidazole? Is there a way to get around
> this?
> Also are there any other tricks to removing backround contaminants
> (under native conditions) apart from use of b-mercaptoethanol and
> higher salt concs?
> I guess that binding and elution parameters are protein specific but I
> would appreciate any guidelines or tips.
> 
> Thanks in advance.
> 
> Cara Baker,
> Dept. of Biochemistry,
> Trinity College Dublin



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