Re: [Histonet] immunohistochemistry and frozen sections
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
> 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 firstname.lastname@example.org
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