Re: In situ with AP and Fast Red TR/Naphthol

From:Robert Schoonhoven


I use BioGenex's New Fuchsin ( which is slightly soluble in
xylene but not soluble in the non-aliphatic hydrocarbons, I use Richard-Allan's
Clear-Rite 3 for clearing instead of xylene when I use this chromogen and
coverslip with Permount.
The New Fuchsin gives a nice bright red end product with Alk. Phos. and is
permanent.   It is also available from Dako and others,  I just happen to like
the BioGenex kit.


Robert Schoonhoven,
Laboratory of Molecular Carcinogenesis & Mutagenesis
University of North Carolina at Chapel Hill

Lisa C Ranford Cartwright wrote:

> Hello histo folk
> We are performing in situ PCR on tissue sections (fixed, embedded,
> sectioned, etc). The PCR results in DIG incorporation into the DNA, which
> we then detect with an anti-DIG antibody conjugated to alkaline
> phosphatase. This part works fine (using a fluorescent detection system).
> We now wish to use a colorimetric substrate rather than a fluorescent one,
> and FAST Red TR/Naphthol had been suggested. This is supposed to give a
> bright red stain. However we also wish to counterstain with Giemsa, and I
> understand the Fast Red is soluble in organics. The final methanol
> concentration in the Giemsa stain will of course be low (5% stain) - is
> this likely to dissolve the fast red deposits?
> I've never used FAst Red TR before. Has anyone tried Fast Red TR and Giemsa
> together? Any thoughts on whether it will work? Any alternatives (note we
> don't want to use NBT/BCIP substrate as our tissue already as some blackish
> material in it which we think will be confused with the signal).
> Thanks for any suggestions, comments and assistance.
> Lisa
> Lisa Ranford-Cartwright,Ph.D.
> Lecturer
> Division of Infection & Immunity
> Institute of Biomedical & Life Sciences
> Joseph Black Building
> University of Glasgow
> Joseph Black Building, Glasgow G12 8QQ
> Scotland, UK
> Tel: 00 44 141 330 2639
> Fax: 00 44 141 330 4600
> Email:
> Lab webpages :
> *****************************************************

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