Figure 2.

Heat-induced antigen retrieval is mandatory to obtain specific nuclear estrogen receptor beta (ERβ) staining. Murine ovaries (A) or uterine tissue (B) were used to evaluate whether antigen unmasking is required for specific nuclear ERβ staining using PPZ0506 antibody. Some sections were stained without antigen retrieval, whereas in others antigen retrieval was performed by heating the sections in sodium citrate buffer (10 mM, 0.05% Tween 20, pH 6.0) in a steamer for 30 min. In the depicted experiment, PPZ0506 antibody was diluted 1:6000. Please note that slides without antigen retrieval showed no nuclear staining at all whereas unspecific cytoplasmatic staining occurs (arrowheads). The used antigen retrieval method enabled a dependable and specific nuclear ERβ staining (of the granulosa cells) in the ovary, but no specific staining in the uterus. Scale bars, 100 µm (in all magnifications).