부산대학교 도서관

Byline: Gregory E. Weitsman (1), George Skliris (1), Kanyarat Ung (1), Baocheng Peng (1), Mamoun Younes (2), Peter H. Watson (3), Leigh C. Murphy (1) Keywords: Estrogen receptor-[beta]; Antibodies; Chromatin immunoprecipitation; Western blotting; Immunohistochemistry Abstract: Several different antibodies to total estrogen receptor (ER)[beta], ER[beta]1 and ER[beta]2/cx have been tested and compared for their ability to immunoprecipitate ER[beta] specific isoforms under chromatin immunoprecipitation conditions (ChIP). The rabbit polyclonal antibodies AP-ER[beta]1 and AP-ER[beta]2/cx, specific for ER[beta]1 and ER[beta]2/cx isoforms, respectively, were the most efficient for ChIP. The monoclonal antibody MCA1974/PPG5/10 was also able to ChIP ER[beta]1, but less efficiently than AP-ER[beta]1. All other antibodies tested were not suitable for ChIP analyses although most antibodies tested immunoprecipitated the appropriate ER[beta] isoforms under standard conditions. To identify antibodies that can also be used to verify in-vivo expression profiles, a comparison of the antibodies to detect ER[beta] isoforms by western blotting and immunohistochemistry was also undertaken. Under the tissue processing and autostaining conditions used at the Manitoba Breast Tumor Bank 385P/GC17, MCA1974/PPG5/10, Ab288/14C8 and MCA2279S/57/3 were found to be the best for IHC of ER[beta] isoforms in human breast tissue biopsy sections, while Ab14021, AP-ER[beta]1 and AP-ER[beta]2/cx were best for western blot detection of ER[beta] isoforms. Author Affiliation: (1) Manitoba Institute of Cell Biology, Department of Biochemistry & Medical Genetics, University of Manitoba, 675 McDermot Avenue, R3E 0V9, Winnipeg, Manitoba, Canada (2) Department of Pathology, Baylor College of Medicine, Houston, Texas, USA (3) Department of Pathology, University of Manitoba, R3E 0V9, Winnipeg, Manitoba, Canada Article History: Registration Date: 12/03/2006 Received Date: 07/03/2006 Accepted Date: 11/03/2006 Online Date: 17/05/2006