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A distinguished team of principal investigators and their associates describe in step-by-step detail a cross-section of the latest research techniques available for studying the endocrine system. As a basis for sophisticated biochemical analysis of receptor properties, the contributors provide methods for the production and purification of a variety of receptors, including progesterone, glucocorticoid, and androgen. Other protocols allow the reader to experiment with DNA binding characteristics, hormone binding assays, and the use of combinatorial chemistry for drug discovery. A series of novel methods utilizing the latest advances in immunochemistry, yeast two-hybrid screening, and fluorescence are included for the detection and analysis of a variety of cellular proteins that influence steroid receptor effectiveness.
Part I. Bioinformatics Bioinformatics of Nuclear Receptors Mark Danielsen Phylogenetic Inference and Parsimony Analysis Llewellyn D. Densmore III Part II. Purification Protocols Expression and Purification of Recombinant Human Progesterone Receptor in Baculovirus and Bacterial Systems Vida Senkus Melvin and Dean P. Edwards High-Yield Purification of Functionally Active Glucocorticoid Receptor Terace M. Fletcher, Barbour S. Warren, Christopher T. Baumann, and Gordon L. Hager Production and Purification of Histidine-Tagged Dihydrotestosterone-Bound Full-Length Human Androgen Receptor Mingmin Liao and Elizabeth M. Wilson Large Scale Production of Nuclear Receptor Ligand-Binding Domains Li-Zhi Mi and Fraydoon Rastinejad Physical Structure of Nuclear Receptor - DNA Complexes Scott A. Chasse and Fraydoon Rastinejad Isolation of Steroid-Regulated Genes from the Uterus by mRNA Differential Display Sushma Kumar, Maarit Angervo, Milan K. Bagchi, and Indrani C. Bagchi Identification of Nuclear Hormone Receptor Homologs by Screening Libraries with Highly Degenerate Oligonucleotide Probes Bruce Blumberg Part III. Steroid Hormone-Binding Assays Use of [99mTc] Technetium-Labeled Steroids as Probes for Steroid Hormone Receptors Frank Wust Steroid Hormone Metabolites and Hormone Binding Assays Rosemary Bland and Martin Hewison Vitamin D3 Analog Screening Sami Vaisanen, Sanna Ryhanen, and Pekka H. Maenpaa Part IV. Protein Interaction Assays Application of Green Fluorescent Protein to the Study of Dynamic Protein - Protein Interactions and Subcellular Trafficking of Steroid Receptors Steven K. Nordeen, Paul R. Housley, Yihong Wan, and Richard N. Day Knockout Mice and Steroid Receptor Research Per Flodby, Stephan Teglund, and Jan-Ake Gustafsson Yeast Two-Hybrid Screening for Proteins that Interact with Nuclear Hormone Receptors Bertrand Le Douarin, David M. Heery, Claudine Gaudon, Elmar vom Baur, and Regine Losson Isolation of a p300/CBP Cointegrator-Associated Protein Coactivator Complex Rabindra N. Bhattacharjee, Caroline Underhill, and Joseph Torchia Nonradioactive Photoaffinity Labeling of Steroid Receptors Using Western Blot Detection System Simon J. Evans and Frank L. Moore Analysis of Steroid Hormone-Induced Histone Acetylation by Chromatin Immunoprecipitation Assay James R. Lambert and Steven K. Nordeen Analyzing the Contributions of Chromatin Structure in Nuclear Hormone Receptor Activated Transcription In Vivo Christy J. Fryer and Trevor K. Archer Cotransfection Assays and Steroid Receptor Biology Shimin Zhang and Mark Danielsen Estrogen Receptor mRNA In Situ Hybridization Using Microprobe System Hironobu Sasano, Sachiko Matsuzaki, and Takashi Suzuki Part V. Cancer Research and Drug Discovery Solid Tumor Cancer Markers and Applications to Steroid Hormone Research Marcia V. Fournier, Katherine J. Martin, and Arthur B. Pardee Assessing Modulation of Estrogenic Activity of Environmental and Pharmaceutical Compounds Using MCF-7 Focus Assay Kathleen F. Arcaro and John F. Gierthy Combinatorial Chemistry in Steroid Receptor Drug Discovery John A. Flygare, Daniel P. Sutherlin, and S. David Brown Identification of Nuclear Receptor Interacting Proteins Using Yeast Two-Hybrid Technology: Applications to Drug Discovery Sunil Nagpal, Corine R. Ghosn, and Roshantha A. S. Chandraratna
This extremely well written and useful collection of laboratory methods for the study of steroid biology is one of a series in Methods in Molecular Biology devoted to steroid hormones and their receptors. The book provides an up-to-date compilation of protocols and assays that are important for the study of steroids and their receptors... it will be invaluable to researchers already in the field to compare protocols and improve methods...The book is divided into sections of bioinformatics, purification protocols, steroid hormone binding assays, protein interaction assays, and cancer research and drug discovery. The bioinformatics section leads off the book with a wealth of information on web sites that are relevant for steroid hormones and their receptors. While there are probably over 40 different proteins involved in steroid hormone receptor interaction, the section on protein interactions is superb. It actually clarifies a good part of the literature as so many of the proteins have different names for the same molecule. Understanding the mechanisms of isolation of these proteins puts the literature into perspective. Furthermore, the protocols are in sufficient detail that our lab was able to adapt several of the techniques immediately. - Doody's Health Sciences Book Review Journal This volume, part of the series Methods in Molecular Biology', describes in detail the protocols and assays used in the study of steroid hormone receptors. - Journal of Pediatric Endocrinology and Metabolism Timely and highly accessible, Steroid Receptor Methods. Protocols and Assays provides biochemists and molecular biologists studying endocrine systems with a full range of the powerful tools needed for elucidating protein interactions with receptors and DNA, as well as for drug discovery. - Biomedicine & Pharmacotherapy Anyone working techniques, or who is tempted to enter the field, would be well advised to obtain this book and read the relevant chapters first. PhD student and established scientist alike will find Steroid Receptor Methods a valuable addition the the library. - Molecular Biotechnology Overall, the book is well written by an impressive list of experts and represents a valuable resource for both inexperienced and seasoned researchers of steroid receptors. - Today's Life Science
