[Histonet] Double stain IHC question

[Gudrun Lang]

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:histonet at lists.utsouthwestern.edu] 
Gesendet: Samstag, 20. Februar 2016 01:54
An: histonet at lists.utsouthwestern.edu
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
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