Q: IF 2nd ABs Mono- or Polyklonal?

Simon Tout s.tout at cclru.unsw.edu.au
Thu Mar 17 04:48:50 EST 1994


In article <2m463l$pe0 at wega.rz.uni-ulm.de>, CCC_GAME at rzmain.rz.uni-ulm.de
(Gamel Anton) wrote:
> 
...stuff deleted...
 
> Now I have to set up a double labelling with two monoklonals
> from mouse differing only in the isotype: IgG1 contra IgG2b.
> There are lots of conjugated polyklonals that discriminate
> IgG1 and IgG2b, what about the cross reactions?
> 
> The experiment should show the two antigens in one cell.
> There are some monoklonal 2nd ABs (only one red) available.
> I suppose  they are much more specific but expensive and all
> from rat. Could there occur other cross reactions e.g.
> Rat at IgG2b binding to a RatIgG2b at MouseIgG1??
> 
> What system should I use monoclonal or polyclonal and are there
> other tricks to get good results in this case?


You should be able to get specific monoclonal- 2nd antibodies for your
different isotypes, just make sure on the technical description that
they're affinity purified and don't cross react. Have you also considered
2nd antibodies to the light chains of your primaries?

Dr.Simon Tout
Co-operative Research Centre for Eye Research and Technology
University of NSW
PO Box 1
Kensington NSW 2033
Australia
s.tout at cclru.unsw.edu.au



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