[Histonet] Immunofluorescence in paraffin sections

Gayle Callis gcallis <@t> montana.edu
Mon Mar 21 11:18:01 CST 2005

Goat is not the only host for seconday antibodies.  You could use Donkey 
antimouse - FITC.   Try Jackson Immunoreasearch for other antibodies that 
will NOT cross react. Jackson has a website.

At 09:21 AM 3/21/2005, you wrote:
>Hello, all,
>I am doing immunofluorescence in paraffin sections. I have some 
>difficulties in it:
>1: For pretreatment of tissue sections, I plan to incubate sections with 
>trypsin solution, but I do not know :what kind of concentration should I 
>choose ? (0.01% trypsin, 0.05% or o.1%)how long does it should last? 
>(10min,20min,30 min)
>2: Recently, I read a protocol about double immunofluorescence, it shows 
>:Antibodies derived from different animal can be mixed and incubated as a 
>cocktail (example: rabbit anti-A and mouse anti-B). The same is valid for 
>secondary antibodies (example: goat anti-rabbit Texas Red conjugated and 
>goat anti-mouse fluorescein conjugated). but If secondary antibodies 
>cross-react, what should I do? My friend told me that I could use 
>depletion method. but I am not sure.
>Can anybody do me a favor?
>Thank you!
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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