anti-His antibody

Bob Steinberg rsteinbe at etowah.ouhsc.edu
Wed Apr 22 17:29:47 EST 1998



Flore Bellefontaine wrote:

> Hi!
> Did someone ever detect with an anti-his antibody a recombinant protein
> purified from pet15B (Novagen)?  The recombinant prot presents a six
> histidines tag at the NH3-term.
> I have been using unsuccesfully the penta-His antibody from Qiagen in
> ELISA, Dot Blot under native and denaturing conditions and in Western Blot
> aswell. Do you have any experience using that antibody?
> Any kind of help is wellcome, thank you in advance,
>
> Flore Bellefontaine
>
> --
> Flore Bellefontaine
> FUNDP
> Unite de Recherche en Biologie Moleculaire (URBM)
> flore.bellefontaine at fundp.ac.be

We've recently obtained a monoclonal anti-H6 antibody from BAbCO (Berkeley
Antibody Company) and found that it reacts with a pET-15b fusion protein in
slot blots. We only have very limited experience so far, but it gave strong
signals at 1/1000, less at 1/4000, and only a relatively weak signal at
1/16,000 (using an alkaline p'tase-conjugated goat anti-mouse antibody and
CDP-Star chemiluminescence for detection). I don't know the sensitivity as of
yet, but it is probably in the low nanogram range.






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