[Histonet] immunofluorescence problem

[slieblich <@t> shaw.ca]

I am doing triple labeling immuno fluorescence of mouse brain tissue and am having some difficulties.  My three markers are NeuN, GFAP, and BrdU.  Previously when I have triple labeled, I only get gfap and brdu fluorescence and no NeuN (or rarely and inconsistently).   I have found out that it is possible that the acid used to denature the DNA for BrdU may be interfering with NeuN antibodies.  I was wondering if anyone has also had this problem with NeuN.  Does anyone have any ideas to get around the NeuN/hydrochloric acid issue?

Thanks!

Stephanie Lieblich
Cellular Neurophysiology Lab
University of British Columbia
Vancouver, B.C., Canada