Re: [Histonet] immunohistochemistry and frozen sections

From:"Greg Dobbin"

You did not mention whether or not the tissues were fixed, so 
assuming you are cutting fresh frozen sections, why are you 
applying atigen retreival methods? Antigen retreival methods are 
only for formalin fixed tissues.

Date sent:      	Tue, 16 Sep 2003 15:48:43 +0100 (BST)
From:           	Vincenzo La Manna 
Subject:        	[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
> _______________________________________________
> Histonet mailing list

Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 University Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851
Making a living is getting; making a life is giving.

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