By Kathleen M. Eyster
This volume examines a big selection of important applied sciences for advancing our realizing of the receptor-mediated activities of estrogen. The protocols during this publication diversity from typical equipment and very important laboratory workhorses, corresponding to receptor binding assays and western blot, to more moderen applied sciences comparable to RNAseq and proximity ligation assay. Chapters additionally speak about protocols from a extensive variety of tissue varieties to illustrate the diversity of estrogen receptor results. Written within the hugely profitable Methods in Molecular Biology series layout, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and tips about troubleshooting and heading off identified pitfalls.
Cutting-edge and thorough, Estrogen Receptors: tools and Protocols is a close and necessary source for scientists who're intrigued by means of the various elements of estrogen.
Read or Download Estrogen Receptors: Methods and Protocols PDF
Similar endocrinology & metabolism books
Obesity and Diabetes (Practical Diabetes)
Style 2 diabetes, linked to weight problems, is this day the most typical type of diabetes. В it's also linked to a couple of different cardiovascular danger elements which represent the metabolic syndrome. В potent administration of diabesity is essential to the relief of morbidity and untimely morbidity as a result of heart problems.
Essential Biochemistry, Endocrinology and Nutrition
Biochemistry is the learn of the chemistry of dwelling organisms, of the ways that meals is used to serve the entire many desires of the physique. Biochemistry is heavily attached with nutrients, the research of the categories and quantities of assorted fabrics required within the nutrition. Biochemistry is usually inextricably int~rtwined with endo crinology, the examine of hormones, for many of the hormones exert their activities through changing the behaviour of chemical reactions in the physique.
- The Official Patient's Sourcebook on Cushing's Syndrome: A Revised and Updated Directory for the Internet Age
- Intermediary Nitrogen Metabolism
- Williams Textbook of Endocrinology, 12e
- Diabétologie
Extra resources for Estrogen Receptors: Methods and Protocols
Example text
4. Binding affinity of membranes kept at −80 °C will probably be reduced over time. 5. Minimum protein content should be above 50 μg and up to 200 μg for a single reaction. 6. 6 nanomolar solution of [3H] 2ME2. 7. Membranes must be soaked for 1 h in binding buffer with BSA prior to use. Acknowledgments The authors would like to acknowledge the laboratory personnel responsible for optimization of this protocol; in particular, the authors wish to acknowledge the graduate student Siva Koganti and fellow Upendra Gumaste.
9. After 1 h, proceed to counting on a Beckman scintillation spectrometer or other suitable platform. 10. Counts (disintegrations per minute, dpm) represent total radioligand binding to the membranes. 4 Competitive Binding for Determining [3H] 2ME2 Specificity 1. In order to determine nonspecific binding of the membranes, preincubate with 1 μM unlabeled 2ME2 in binding buffer for 10 min before the addition of the [3H] 2ME2 radioligand. 2. 3. 3. Specific [3H] 2ME2 binding is defined as that portion of the total binding displaced by unlabeled 2ME2.
Combining transcriptomic and ChIP-seq data enables a distinction to be drawn between direct and indirect estrogen target genes. This chapter discusses some methods of identifying estrogen target genes that do not require any expertise in programming languages or complex bioinformatics. Key words Estrogen, ChIP-seq, Transcriptomics, Gene targets, Bioinformatics 1 Introduction Gender disparities are associated with the risk of multiple diseases [1]. Estrogen is clearly associated with the risk of many gynecological malignancies but also has a role in modulating aspects of autoimmunity [2–4].