Re: fluorescent peroxidase substrate
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of histonet) |
To: | histonet@histosearch.com |
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On Thu, 2 Dec 1999, Corazon Bucana wrote:
> Just out of curiousity, is there a peroxidase substrate for
> immunohistochemistry that forms an insoluble precipitate and at the same
> time is also fluorescent?
Yes. The procedure known as TAT (tyramide amplification
technique) or CARD (catalysed reporter deposition) is a
sensitive method for peroxidase. It generates a fluorescent
product that is covalently bound to the tissue at sites
of enzymatic activity. The chromogen is a compound made by
linking tyramine to a fluorochrome. In the presence of
peroxidase and its substrate, hydrogen peroxide, the
tyramine half of the chromogen molecule is oxidized to
a highly reactive free radical, which immediately reacts
with and binds to protein (probably mainly to tyrosine)
in the tissue. Simple methods for synthesizing fluorescently
labelled tyramides are described by Hopman,AHN et al (1998)
in J Histochem Cytochem 46:771-777.
I haven't tried this myself, but the published pictures
are impressive. It is used more for in situ hybridization
than for imunohistochemistry, but is suitable for any
technique in which peroxidase activity needs to be
localized.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
E-mail: kiernan@uwo.ca
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