imidazol eluent alternative in his-tagged proteins

Soenke Behrends behrends at plexus.uke.uni-hamburg.de
Sat Oct 30 09:56:18 EST 1999


Dear netters,

I am working on a hemoprotein where the heme is bound
to the protein by a histidin (and specifically by the imidazol
moiety). My impression and fear is, that after his-tag binding
to a Ni column the elution with imidazol buffer not only 
destroys the protein column interaction but also frees 
my protein of heme. Does anyone know of an alternative
way of elution (under native conditions) get the protein
off the column. 

I have thought also to construct a  His-tag with TEV protease
site and cleave it off the column, but I am forced to tag
the C-terminus and I have not found any vector / technique
to do that. 

Thanks a lot for any hint or comment
Soenke







More information about the Methods mailing list