I am trying to stain mouse brain sections (coronal) for B-catenin and look for nuclear localization of B-catenin. The brains were PFA-perfised and stained using the free-floating section method. I used Sigma Antibody (rabbit-whole antiserum C2206) at 1:1000 and 1:500 in PBS + 10% donkey serum (same species as the secondary) + 0.1% Triton-X100. However, I have a high background and it is difficult to tell if the nuclear staining is genuine or is due to the background. Also, the staining is not consistent across sections (stained in the same wells- receiving the same treatment), so I would sometimes need to change the scanning parameters (confocal microscope) to get close enough signal in different images, which confuses matters even more. Does anyone have experience with B-catenin staining or know of a good antibody?