Do IHC sequentially. The first one is with a permanent chromogen (DAB,
BCIP), then place slides in a chaotrophic buffer at elevated temperature (we
microwave slides at 10sec/slide in a Retrievit-8 buffer). Wash and proceed
with the second IHC, starting from the protein blocking step.
Abizar
www.innogene.com



> From: Sunil Thomas K [mailto:sthomaska@yahoo.com]
> Sent: Monday, July 23, 2001 6:25 AM
> To: Bruce Abaloz
> Cc: histonet@pathology.swmed.edu
> Subject: Re: IHC Double staining
>
>
>
>
>
> --- Bruce Abaloz <b.abaloz@zoology.unimelb.edu.au>
> wrote:
>
>  can not find a protocol or a kit that would
> > allow me to achieve this with two antibodies raised
> > in the same species.
>
> I am also a student-my opinion is that, if you use
> primary antibodies raised in same species how will you
> know what the secondary antibody is recognising.
> The alternative is to use  primary antibodies that are
> tagged to different labels. the specificity of such
> detection will be low.
> Sunil Thomas K
>
> > Histonetters,
> >
> > I am a student in the department of Zoology at the
> > University of Melbourne, Australia.  I have two