RE: Direct FA on FFPE

From:"Greer, Patricia"

You should not have a problem using the FA conjugates on FFPE sections. We
use a couple of antibodies that are only available as FITC conjugates. We
incubate the sections with the conjugate as we do our unconjugated primary
antibodies, followed by incubation with mouse anti- FITC (Dako M0878).  Then
follow our same protocol for detection - we use biotinylated X-mouseX-rabbit
link then strep avidin-alkaline phosphatase followed by fast red substrate
(Dako's LSAB2 kit).  Works great for us - just remember you will need to
titrate the anti-FITC as well as the primary (FA conjugate) for optimum

Pat Greer
Atlanta, GA

Hello All,

I have a question about using direct fluorescent antibody staining 
on formalin-fixed paraffin embedded (FFPE) tissue sections. 

Can these antibodies be used reliably for FFPE sections? In an 
earlier post (subject: Veterinary Viral Antibodies), I mentioned that 
the only source I could find of 2 viral antiseras was VMRD and they 
only sold the Direct Fluorescent Conjugated form, but I am not sure 
these will work. Any thoughts from the "Immuno-Gurus" out there? 
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
"A farmer is a person outstanding in their field."

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