RE: Direct FA on FFPE
Greg,
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
dilutions.
Pat Greer
CDC/IDPA
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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