Re: converting a chromogen signal to fluorescence

From:Mikael Niku

Philopena, Jennifer wrote:
> Hi All.
> I have an HRP-conjugated primary antibody that works very well for IHC (and
> I'd like to keep using it), but I want a fluorescent signal.  Does anyone
> know of a fluor-conjugated anti-HRP antibody?  

A solution would be to use a fluorochrome-conjugated tyramide
reagent. The peroxidase converts the tyramide into
a radical that rapidly forms covalent bonds with adjacent
proteins, meaning that in effect you get lots of
fluorochrome bound around the antigen (don't get me
wrong, the spatial resolution is still OK).

This is easy (after HRP-antibody you just add the tyramide
reagent, incubate 10 min or so, wash, and that's it) and
should produce pretty strong signal (I have myself used
only biotinylated tyramide, but the principle is
the same) . The only drawback I can think of is that
the stuff is pretty expensive. It's patented by NEN
and sold also by a few other companies (I think DAKO
and perhaps Molecular Probes).

    Mikael Niku             URL:
    University of Helsinki  Dept. Basic Veterinary Sciences
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