I have read that this is possible using the extremely
sensitive tyramide signal amplification (TSA) system.
The idea is that you first stain with one antibody
at such a high dilution that it can only be visualized
by TSA. Then you do a conventional stain with the other.

I haven't done this myself, but it has been described at

I think I've seen some publications also, so you could
try a Medline search.

You'll find information on the TSA system itself at

   Mikael Niku             URL:
   University of Helsinki  Dept. Basic Veterinary Sciences
       - Mit=E4k=F6 mielt=E4 olen l=E4nsimaisesta sivistyksest=E4?
         Minusta se olisi erinomainen ajatus!
                                              - Gandhi
----- Original Message -----
From: "Bruce Abaloz" <>
To: <>
Sent: Thursday, July 19, 2001 3:33 AM
Subject: IHC Double staining

> Histonetters,
> I am a student in the department of Zoology at the University of
Melbourne, Australia.  I have two antibodies raised in the same species
however I need to co-localize their  expression in the one tissue section.
I have protocols for immuno double staining of antibodies raised in
different species, but I can not find a protocol or a kit that would allo=
me to achieve this with two antibodies raised in the same species.  I am
currently using immunofluorescence and would like to remain with this
visualization.  I would be very grateful for any help.
> Thanks
> Doug


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