Estrogen Receptors: Methods and Protocols

ISBN-13: 9781071619193 / Angielski / Twarda / 2022

Kathleen M. Eyster

Spis treści

1. Antibody Validation for Estrogen Receptor Beta

            Madeleine Birgersson, Borbala Katona, Cecilia Lindskog Bergström, Fredrik Pontén, and Cecilia Williams

 

2. Immunoblot Detection of the Phosphorylation of the Estrogen Receptor a as an Outcome of GPR30/GPER1 Activation

            Sara Clark, Kevin Pollard, Jennifer Rainville, and Nandini Vasudevan

 

3. Immunoprecipitation Analyses of Estrogen Receptor α Phosphorylated at Serine 216 in the Mouse Liver

            MyeongJin Yi, Sawako Shindo, and Masahiko Negishi

 

4. Immunofluorescent Verification of Silencing Estrogen Receptor α with siRNA in the Intact Rodent Brain

            Ana C. Ribeiro, Mihaela A. Stavarache, Sergei Musatov, and Donald W. Pfaff

 

5. Detection and Functional Analysis of Estrogen Receptor α Phosphorylated at Serine 216 in Mouse Neutrophils

            Sawako Shindo, Rick Moore, MyeongJin Yi, and Masahiko Negishi

 

6. Quantification of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate in Response to Estrogen

            Thomas W. Bonagura, Jeffery S. Babischkin, Gerald J. Pepe, and Eugene D. Albrecht

 

7. In Situ Hybridization of Estrogen Receptors α and β in the Human Testis

            Katja Hartmann and Daniela Fietz

 

8. Extraction of RNA and Analysis of Estrogen-Responsive Genes by RT-qPCR

            Michelle L. Booze and Kathleen M. Eyster

 

9. PET Imaging of Estrogen Receptors Using ¹⁸F-Based Radioligands

            Manoj Kumar, Kelley Salem, Justin J. Jeffery, and Amy M. Fowler

 

10. The Use of ERE-Luc Reporter Mice to Monitor Estrogen Receptor Transcriptional Activity in a Spatio-Temporal Dimension

            Sara Della Torre, Elisabetta Vegeto, and Paolo Ciana

 

11. Use of Reporter Genes to Analyze Estrogen Response: The Transgenic Zebrafish Model

            Daniel A. Gorelick, Caroline Lucia Pinto, Ruixin Hao, Silvia Karim, and Maria Bondesson

 

12. Assessing Estrogenic Activity of Classical Estrogen Receptor‑Binding Compounds

            Richard A. Pepermans and Eric R. Prossnitz

 

13. An Optimized ChIP-Seq Protocol to Determine Chromatin Binding of Estrogen Receptor Beta

            Rajitha Indukuri, Anastasios Damdimopoulos, and Cecilia Williams

 

14. Polysome-Fractionation for Transcriptome-Wide Studies of mRNA Translation

            Johannes Ristau, Kathleen Watt, Christian Oertlin, and Ola Larsson

 

15. Anota2seq Analysis for Transcriptome-Wide Studies of mRNA Translation

            Christian Oertlin, Kathleen Watt, Johannes Ristau, and Ola Larsson

 

16. Whole-Genome Genotyping Using DNA Microarrays for Population Genetics

            Austin J. Van Asselt and Erik A. Ehli

 

17. Proteomics Analysis of the Estrogen Effects in the Rat Uterus Using Gel-LC and Tandem Mass Spectrometry Approaches

            Maria D. Paez and Eduardo A. Callegari

 

18. Expression Profiles of Estrogen-Regulated MicroRNAs in Cancer Cells

            Amena Archer, Claudia Kutter, and Cecilia Williams

 

19. Molecular Cloning and Purification of the Protein Lysine Methyltransferase SMYD2 and Its Co-Crystallization with a Target Peptide from Estrogen Receptor Alpha

            Yingxue Zhang and Zhe Yang

 

20. Molecular Design, Synthesis, and Evaluation of SNIPER(ER) that Induces Targeted Protein Degradation of ERα

            Nobumichi Ohoka, Hidetomo Yokoo, Keiichiro Okuhira, Yosuke Demizu, and Mikihiko Naito

 

21. Assessment of Metabolic Regulation by Estrogen Receptors

            Geetanjali Sharma and Eric R. Prossnitz

 

22. RNA-Seq Experiment and Data Analysis

            Miyuraj Harishchandra Hikkaduwa Withanage, Hanquan Liang, and Erliang Zeng

Blurb

This fully updated collection includes chapters on a wide array of techniques that are vital for advancing our understanding of the physiological and pathological effects of estrogen, mediated by estrogen receptor α (ERα) and estrogen receptor β (ERβ) as well as the non-genomic membrane-bound G-protein coupled estrogen receptor (GPER/GPR30). The protocols range from standard methods and important laboratory workhorses such as immunoblot and RT-qPCR, to newer technologies such as PET imaging, reporter animal models, polysome fractionation, and many others, involving a broad range of tissue types to demonstrate the variety of estrogen receptor effects. Written for this highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Estrogen Receptors: Methods and Protocols, Second Edition serves as an ideal guide to researchers working to explore the vibrancy and excitement of the field of estrogen receptors and estrogen actions.