1. An Optimized ChIP-Seq Protocol to Determine Chromatin Binding of Estrogen Receptor Beta.
- Author
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Indukuri R, Damdimopoulos A, and Williams C
- Subjects
- Chromatin genetics, Chromatin Immunoprecipitation methods, Estrogens, Chromatin Immunoprecipitation Sequencing, Estrogen Receptor beta genetics, Estrogen Receptor beta metabolism
- Abstract
Estrogen regulates transcription through two nuclear receptors, ERα and ERβ, in a tissue and cellular-dependent manner. Both the receptors bind estrogen and activate transcription through direct or indirect interactions with DNA. Revealing their interactions with the chromatin is key to understanding their transcriptional activities and their biological functions. Chromatin-immunoprecipitation followed by sequencing (ChIP-Seq) is a powerful technique to map protein-DNA interactions at precise genomic locations. The genome-wide binding of ERα has been extensively studied. Similar studies of ERβ, however, have been more difficult, in part due to a lack of endogenous expression in cell lines and lack of specific antibodies. In this chapter, we provide an optimized stepwise ChIP protocol for a well-validated ERβ antibody, which is applicable for ChIP-Seq analysis of cell lines with exogenous expression of ERβ., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2022
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