histidine tagging

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Tue Oct 29 20:19:52 EST 2002


"Nora Plesofsky" <nora at biosci.cbs.umn.edu> wrote in message
news:8DF774B2-EB7B-11D6-97A0-00306577FE10 at biosci.cbs.umn.edu...
> I wish to tag proteins that I have engineered to be expressed in my
> organism. It seems to me that hexahistidine tagging would serve two
> purposes, i.e. allowing visualization of the protein by immunoblotting
> and also affinity isolation of the protein with attendant proteins. I
> am asking if others have had experience with antibody reactivity to the
> histidine tag and with background reactivity. Is there another tag that
> has served this purpose possibly better?

His-tag antibodies worked well for us. There is also a fluorescent His-tag
detection system which sometimes works and sometimes does not. We have also
had good immunoblotting results with pretty much all kinds of fusions and
tags that we tried - MBP, GST, thioredoxin, His6, Biotin, FLAG, Strep,
chitin affinity, and some others. Without addressing your specific problem,
it's hard to say which option will work better.

A.G.E.





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