In article <4hm2ko$5u3 at kralle.zdv.Uni-Mainz.DE>, Tanja
<jschroed at vzdmza.zdv.uni-mainz.de> wrote:
> I am quite new to handle with Immunoaffinity Chromatography and I want
to purify a protein from Hordeum vulgare with this subject.
> Before I start to purify the protein. I have to clean the polyclonal
mouse IgG with a protein A column (BioRad, Affi-Gel Protein A G=
> el). The antibody did not bound to the protein A and I found the
complete IgG in the first wash step. Then I tried a different way:
> I tried to couple the IgG via caboanhydrate moities to hydrazide
activated agarose beats (BioRad, Affi-Gel Hz Immunoaffinity Column)=
> In the first step I detected again the complete IgG so it did not bind
to the gel.
> Maybe there are eome hints behind the productinformations?
Dear Tanja,
if your Protein A column hasn't worked in the first place, why did
you immediately changed the technique ? Protein A on polyclonal mouse
serum simply must give you something; otherwise some more basic is
^^^^^^
totally wrong. But to help you, one would require some more information,
how you detect the IgG, what do you understand under 'I detected again
the complete IgG', ...
^^^^^^^^^^
BTW, a verg good textbook for antibodies is Goding, Monoclonal Antibodies:
principles and practice, 2nd ed., San Diego, CA, USA, Academic Press 1986.
In addition, you just might ask some immunologist's in Mainz, in the
group of Prof. Ruede most of the people should have used Protein A columns.
Good luck,
jorg
kirberg at nki.nl
