mrc7 at cam.ac.uk
Tue Oct 1 06:23:10 EST 2002
In article <66becbd5.0209301205.5a01b2f7 at posting.google.com>, Daniel Rosen
<URL:mailto:daffyduck1945 at hotmail.com> wrote:
> This is probably a very basic question, but what could cause isotype
> cross-reactivity in antibodies? I'm trying to raise rabbit anti-mouse
> IgG antibodies by injecting the rabbit with whole mouse IgG (H&L), but
> find that the antibodies are also reactive to the other isotypes.
> Is this because I'm using whole mouse IgG and the antibody recognizes
> the light chains of other antibodies?
Almost certainly yes. You also have V-region epitopes which would be shared
with other classes and subclasses.
> What should I do instead? Use only the gamma chain?
That is one method but you might not get specificity for native epitopes.
> Is there a way to extract just the IgG antibodies?
Yes the method is to pre-adsorb your antisera on the isotypes you don't
want to detect. You could also combine this with affinity purification on
> Thanks to all who reply!
Mike Clark, <URL:http://www.path.cam.ac.uk/~mrc7/>
o/ \\ // || ,_ o M.R. Clark, PhD. Division of Immunology
<\__,\\ // __o || / /\, Cambridge University, Dept. Pathology
"> || _`\<,_ // \\ \> | Tennis Court Rd., Cambridge CB2 1QP
` || (_)/ (_) // \\ \_ Tel.+44 1223 333705 Fax.+44 1223 333875
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