Isotype Cross-Reactivity

[Mike Clark]

In article <66becbd5.0209301205.5a01b2f7 at posting.google.com>, Daniel Rosen
<URL:mailto:daffyduck1945 at hotmail.com> wrote:
> Hi,
> 
> 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
IgG.

> Thanks to all who reply!
> 
> 
> Dan
> 


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