Development and Characterization of Novel Rat Anti-mER beta Sera

作者：Novaira Horacio J^*; Graceli J B; Capellino S; Schoeffield A; Hoffman G E; Wolfe A; Wondisford F; Radovick S

来源：Endocrinology, 2016, 157(7): 2844-2852.

DOI：10.1210/en.2016-1122

Estrogens regulate normal sexual and reproductive development in females. Their actions are mediated mainly by estrogen receptor (ER)alpha and ER beta. Understanding the function of ERs necessitates knowing their cellular location and protein partners, which, in turn, requires reliable and specific antibodies. Several antibodies are available for ER alpha; however, discrepancies in immunoreactivity have been reported for ER beta. Here, we have developed antisera for mouse ER beta (mER beta) using a specific C-terminal 18-amino acid peptide conjugated to mariculture keyhole limpet hemocyanin. Sprague Dawley rats were immunized, and the resulting antisera were characterized by Western blot analysis of nuclear extracts from tissues of wild-type (WT) mice, and mice genetically modified to lack either ER alpha (CER alpha KO) or ER beta (CER beta KO). An approximately 56-kDa protein was detected in the hypothalamus, uterus, ovary, mammary gland, testes, and epididymis of WT mice, consistent with the predicted molecular size of ER beta. In addition, the same protein band was identified in in vitro synthesized mER beta protein and in the mammary glands of CER alpha KO mice. The approximately 56-kDa protein was not observed in in vitro synthesized mER alpha protein or in any tissue examined in the CER beta KO mice. Immunohistochemistry using the antisera revealed ER beta staining in the granulosa cells of WT ovaries and in the mediobasal hypothalamus, paraventricular nucleus, and cerebral cortex in the WT adult mouse brain. These data suggest that the novel rat anti-mER beta sera are specific to ER beta to allow investigators to explore to cellular and physiological role of ER beta in the brain and other mouse tissues.

• 出版日期2016-7
• 单位rutgers