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Antibody Validation for Estrogen Receptor Beta.
Birgersson, Madeleine; Katona, Borbala; Lindskog, Cecilia; Pontén, Fredrik; Williams, Cecilia.
Afiliação
  • Birgersson M; Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
  • Katona B; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
  • Lindskog C; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
  • Pontén F; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
  • Williams C; Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden. cecilia.williams@scilifelab.se.
Methods Mol Biol ; 2418: 1-23, 2022.
Article em En | MEDLINE | ID: mdl-35119656
Antibodies can cross-react with proteins other than their intended targets, and antibody-based applications can, if not properly validated, lead to flawed interpretations. When evaluating 13 anti-estrogen receptor beta (ERß) antibodies in 2017, we concluded that only one of them was specific. Applying this antibody in immunohistochemistry of over 44 different normal human tissues and 20 types of cancers revealed ERß expression in only a few selected tissues. This aligned with mRNA evidence but contradicted a large set of published literature. ERß protein expression continues to be reported in tissues without clear support by mRNA expression. In this chapter, we describe how ERß antibodies can be thoroughly validated and discuss selection of well-characterized positive and negative controls. The validation scheme presented is applicable for immunohistochemistry and Western blotting. The protocol includes evaluation of mRNA evidence, use of public databases, assessment of on- and off-target binding, and an optional step for corroboration with immunoprecipitation and mass spectrometry.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Receptor beta de Estrogênio / Anticorpos Limite: Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Suécia

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Receptor beta de Estrogênio / Anticorpos Limite: Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Suécia