TY - JOUR
T1 - Development and characterization of novel rat anti-mERβ sera
AU - Novaira, Horacio J.
AU - Graceli, J. B.
AU - Capellino, S.
AU - Schoeffield, A.
AU - Hoffman, G. E.
AU - Wolfe, A.
AU - Wondisford, F.
AU - Radovick, S.
N1 - Funding Information:
This work was supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development/National Institutes of Health R01 HD068777-01A1, and through cooperative agreement (partnership U01 HD066432-01) as part of the Cooperative Partnerships to Promote Workforce Diversity in the Reproductive Sciences Program; and the Ciencias Sem Fronteiras scholarship grant from Coordenacao de Aperfeicoamento de Pessoal de Nível Superior (CAPES)/Brazil (1819-61-2)
Publisher Copyright:
Copyright © 2016 by the Endocrine Society.
PY - 2016/7
Y1 - 2016/7
N2 - Estrogens regulate normal sexual and reproductive development in females. Their actions are mediated mainly by estrogen receptor (ER)α and ERβ. 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 ERα however, discrepancies in immunore activity have been reported for ERβ. Here, we have developed antisera for mouse ERβ (mERβ) 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 ERα (CERαKO) or ERβ (CERβKO). 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 ERβ. In addition, the same protein band was identified in in vitro synthesized mERα protein and in the mammary glands of CERαKO mice. The approximately 56-kDa protein was not observed in in vitro synthesized mERβ protein or in any tissue examined in the CERβKOmice. Immunohisto chemistry using the antisera revealed ERβ 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-mERβ sera are specific to ERβ to allow investigators to explore to cellularandphysiological role of ERβ in the brain and other mouse tissues.
AB - Estrogens regulate normal sexual and reproductive development in females. Their actions are mediated mainly by estrogen receptor (ER)α and ERβ. 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 ERα however, discrepancies in immunore activity have been reported for ERβ. Here, we have developed antisera for mouse ERβ (mERβ) 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 ERα (CERαKO) or ERβ (CERβKO). 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 ERβ. In addition, the same protein band was identified in in vitro synthesized mERα protein and in the mammary glands of CERαKO mice. The approximately 56-kDa protein was not observed in in vitro synthesized mERβ protein or in any tissue examined in the CERβKOmice. Immunohisto chemistry using the antisera revealed ERβ 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-mERβ sera are specific to ERβ to allow investigators to explore to cellularandphysiological role of ERβ in the brain and other mouse tissues.
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U2 - 10.1210/en.2016-1122
DO - 10.1210/en.2016-1122
M3 - Article
C2 - 27105387
AN - SCOPUS:84977123308
SN - 0013-7227
VL - 157
SP - 2844
EP - 2852
JO - Endocrinology
JF - Endocrinology
IS - 7
ER -