I stumbled into immunofluorescence for the first time and could use some
advice. I am trying to stain GFP on formalin fixed paraffin embedded
sections. I have a conjugated alexa fluor 488 anti-gfp antibody from
invitrogen that I've now found out was not tested on paraffin sections.
I have seen articles supporting and denying that it works. In addition,
do I retrieve or not as again, I've seen literature supporting both.
Moreover, one article cut sections at 12 microns. My protocol for my
first run consisted of a protein block for 10 minutes, blowing off,
1:400 of the conjugated alexa fluor 488 ant-gfp antibody for 1 hour at
room temp., buffer rinse, DI water rinse, aqueous mounting medium, and
coverslip. To my best ability I performed everything in the dark. The
results were that I had no fluorescing whatsoever!! Any help would be
MaryAnn Dixon BS
UF Veterinary Medical Center
(352) 392-2235 Ext. 4517
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