[Protein-analysis] Re: good hexa-his antibody supplier wanted

allisonh via proteins%40net.bio.net (by allison from nospam.com)
Thu Oct 4 12:52:04 EST 2007


Jurre Hageman wrote:
> Hi all,
> 
> I am working with 6XHis-tagged proteins for Ninta pulldown assays. I
> subcloned a library of genes in this His vector that I generated by
> oligocloning of  Met-6Xhis  encoding oligos. So, the His is at the
> N-terminus and just 6XHis. Unfortunately, I have severe troubles to detect
> my proteins with either a penta-his AB (qiagen) or a AB against 6XHis (Santa
> Cruz, 16-002). I know that the cloning and expression is OK as the pulldown
> works fine and I can detect the protein by the AB against the protein. but
> for most proteins in my library i do not have a specific AB.
> 
> It seems that most people generated a larger tag that contains some
> additional amino acids to generate a stronger epitope. We have a good his ab
> that recognizes c-terminal his-fusions (it requires a his-cooh) but this is
> not functional for this library of course.
> 
> Is there anyone out there that knows of a good working 6XHis AB which also
> recognizes plain 6XHis fusions at the N-terminus of the protein of interest.
> 
> Thanks!!!!
> 
> 
> Jurre


I have used anti-His from Qiagen.  They have 3 anti His antibodies, 
maybe one would work for you:
http://www1.qiagen.com/Products/Protein/Detection/QIAexpressDetectionSystem/Anti-HisAntibodySelectorKit.aspx?rp=1000018&rpg=0&ShowInfo=1

NAYY
Allison


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