In my opinion, this would only be possible, if the commercial and the homegrown antibody are from different species. For example one from mouse and one from rabbit. Then you can proceed with different secondaries (goat anti mouse conjugated with peroxidase, goat anti rabbit conjugated with alkaline phosphatase). Then chromogens that work with each of the enzymes.
If the antibodies are from the same species I see no way to distinguish both. Only if one is conjugated with biotin and the other with digoxigenin, then you could proceed with secondaries against biotin and digoxigenin. etc.. Gudrun -----Ursprüngliche Nachricht----- Von: Judi Ford via Histonet [mailto:[email protected]] Gesendet: Samstag, 20. Februar 2016 01:54 An: [email protected] Betreff: [Histonet] Double stain IHC question Hi everyone, I have a question in chromogenic double staining. Here is the situation. Tissue = human, frozen Antibody = same protein (A) 1. Commercial antibody of A 2. Homegrown antibody of A, human, biotinylated Question: can you stain both versions of this antibody on the same tissue, same slide? Goal is to see where each stains in the tissue and if they co-localize. If they do co-localize then how do you distinguish between that and where they stain individually? Would you use different chromogens and hope that where they come together it turns a different color? I am really interested if this can work. Thanks in advance for any replies. Judi South San Francisco, CA _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
