Re: [Histonet] immunohistochemistry and frozen sections

If it's not an inpolite question, why are you trying to demonstrate estrogen receptors on frozen tissue?
Is this absolutely neccessary as estrogen receptors are very easily demonstrated on paraffin processed tissue, which maintains better morphology and gets almost guaranteed results.

We use the DAKO 1D5 clone on FFPE tissue using Vector Unmasking fluid for Antigen Retrieval and the DAKO ChemMate detection system and get excellent results every time.

Nick Kirk
Hinchingbrooke Hospital

>  from:    Vincenzo La Manna 
>  date:    Tue, 16 Sep 2003 15:48:43
>  to:
>  subject: Re: [Histonet] immunohistochemistry and frozen sections
> Hi Histonetters,
> I've been struggling for 4 months on my 5 microns
> frozen sections trying to find estrogen receptors.
> I've tried both immunoperoxidase kit from Vector
> laboratories either indirect immunofluorecsence (FITC
> Goat anti mouse Secondary Ab from Sigma) but I've not
> yet obtained good results.
> My primary Ab is a mouse monoclonal to bovine
> Estrogen receptor from Cymbus biotechnology.
> Sections come from tissue that is supposed to be a
> non-target tissue (mid laminar region from cow's
> hooves)blocking with 2% goat normal serum (sigma)
> whash in tbs buffer
> All different methods for antigen retrieval have been
> tested but sections look poor regarding histologic
> quality and I still have very big problems with non
> specific binding an staining (indirect
> immunofluorescence).
> It looks like frozen sections are too tender to
> efford the whole processing.
> Every suggestion or reference is welcome,
> thank U
> La Manna Vincenzo
> Department of Agriculture and Forestry, University of
> Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK.
> Telephone; 01224 274259
> Fax; 01224 273731
> e-mail
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