[Histonet] anti-GFP

[MaryAnn Dixon]

Hi histonetters,

 

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
appreciated.  

 

MaryAnn Dixon  BS

Biological Scientist

Anatomic Pathology

UF Veterinary Medical Center

(352) 392-2235 Ext. 4517