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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet