Anti erβ antibody

Prostate ventral lobes and the bladder were harvested for histological analyses after cystometry. One part of the prostate and the bladder was fixed in buffered 10% formaldehyde solution for 24 hr, embedded in paraffin, cut with a microtome, and stained with hematoxylin-eosin for evaluating tissue inflammation. The remainder of the prostate was used for immunohistochemical staining. Paraffin embedded prostate sections were placed on silicone-coated slides. After deparaffinization in xylene and rehydration using graded alcohol solutions, the sections were fixed with 10 mM sodium citrate pH 6 at 105°C by Autoclave (TOMY SEIKO co. Ltd.) for antigen retrieval. The tissues were rinsed with phosphate buffered saline (PBS), transferred to 0.3% hydrogen peroxide for 10 min to block peroxidase activity, and rinsed with distilled water and PBS. The sections were blocked with 10% normal goat serum (NICHIREI CORPORATION.) for 30 min. These tissues were incubated with an anti-ERβ antibody (1:1,000; Santa Cruz Biotechnology, Inc.) for 1 hr at room temperature. After washing with PBS, the tissue sections were incubated for 30 min with HRP-labeled polymer anti-rabbit antibody (DAKO.) at room temperature. After washing with PBS, the color was developed using the Dako Cytomation Liquid DAB Substrate Chromogen System (DAKO). The tissues were counterstained with hematoxylin.