His tagged protein won't bind

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Thu Sep 23 05:35:15 EST 2004

"Dr. Barry Panaretou" wrote:

> We've tagged a protein with 6xHis, and expressed it in E.coli. It didn't
> bind nickel resin. We also tagged the C -terminus and again it didn't
> bond to nickel resin. Has anyone come across this before (protein is in
> supernatant after hard spin, so seems soluble--can also detrect with
> anti-his antobody on westerns, in both cases)

Sometimes tags get hidden inside the folded protein. Two options: 
a) classical protein purification not relying on the tag 
b) denature the protein to make the tag available

More information about the Proteins mailing list

Send comments to us at biosci-help [At] net.bio.net