his tags

Yoram Gerchman yoramg2 at LEONARDO.LS.HUJI.AC.IL
Fri Oct 31 07:18:57 EST 1997


> ----------
> From:
> burdett at ABACUS.MC.DUKE.EDU[SMTP:burdett at ABACUS.MC.DUKE.EDU]
> Sent: Friday 24 de October de 1997 17:14
> To: methods at net.bio.net
> Subject: his tags
> 
> I have been having a lot of trouble getting my his-tagged protein to
> bind well to any metal affinity column and see about 70% of the
> material
> in the 'pass-thru' fraction. My protein is poorly made under any
> condition and there is not a lot of material in inclusion bodies for
> example. Are there any suggestions out on the net?
> 
I had this problem once trying to purify from chloroplast. If I remember
correct we ended up pre-purifieng on a DEAE column, apperently something
in the extract was disturbing the binding. We use Qiagen NTA-Ni.
.........................................................................
Yoram Gerchman
Division of Microbial & Molecular Ecology
The Institute of Life Sciences
Hebrew University of Jerusalem
Givat Ram, Jerusalem 91904, Israel
Tel: 972-2-6585175




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