This volume presents valuable techniques for studying the class of ligand-activated transcription factors known as nuclear receptors. After a brief overview of the history of the field, chapters cover methods to detect the receptors and their mRNAs in various tissues, protocols to characterize nuclear receptor modulators and activities, their signaling and roles in certain pathogenesis, molecular modeling of nuclear receptor-ligand interactions, as well as the utility of informatics in the field of nuclear receptors. Written in the highly successful Methods in Molecular Biology series format,…mehr
This volume presents valuable techniques for studying the class of ligand-activated transcription factors known as nuclear receptors. After a brief overview of the history of the field, chapters cover methods to detect the receptors and their mRNAs in various tissues, protocols to characterize nuclear receptor modulators and activities, their signaling and roles in certain pathogenesis, molecular modeling of nuclear receptor-ligand interactions, as well as the utility of informatics in the field of nuclear receptors. Written in the highly successful Methods in Molecular Biology series format, 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 cutting-edge, Nuclear Receptors: Methods and Experimental Protocols seeks to aid researchers working toward furthering ourunderstanding of these vital receptors and their role in numerous pathological conditions.
Nuclear Receptors: A Historical Perspective.- An Optimized Immunoblotting Protocol for Accurate Detection of Endogenous PGC-1 Isoforms in Various Rodent Tissues.- Sub-Cellular Localization of NR4A2 Orphan Nuclear Receptor Expression in Human and Mouse Synovial Joint Tissue.- In Vivo Quantitation of Estrogen Receptor b Subtype Expression in Ovarian Surface Epithelium Using Immunofluorescence Profiling and Confocal Microscopy.- In VivoChIP-Seq of Nuclear Receptors: A Rough Guide to Transform Frozen Tissues into High Confidence Genome-Wide Binding Profiles.- High-Content Analysis of Constitutive Androstane Receptor Nuclear Translocation.- Antibody Validation Strategy for Nuclear Receptors.- Immunofluorescence Labeling of Nuclear Receptor Expression in Formalin-Fixed Paraffin-Embedded Tissue.- Detection of ADP-Ribosylation of the Androgen Receptor Using the Recombinant Macrodomain AF1521 from Archaeoglobus fulgidus.- Reconstitution of the Steroid ReceptorHeterocomplex.- High-Throughput Imaging of PPIX Using Confocal Microscopy.- PGC-1 Overexpression via Local In Vivo Transfection in Mouse Skeletal Muscle.- A Reverse Transfection Method for Screening of Nuclear Receptor Activators.- Hybrid Reporter Gene Assays: Versatile In Vitro Tools to Characterize Nuclear Receptor Modulators.- Analysis of the Transcriptional Activity of Retinoic Acid-Related Orphan Receptors (RORs) and Inhibition by Inverse Agonists.- Examining the Role of Nuclear Receptors during In Vivo Chemical-Mediated Breast Tumorigenesis.- Analysis of IL-4/STAT6 Signaling in Macrophages.- Isolation and Characterization of Adipose Tissue Macrophages.- Assessing Mitochondrial Bioenergetics in Isolated Mitochondria from Mouse Heart Tissues Using Oroboros 2k-Oxygraph.- Generation of Skeletal Myocytes from Embryonic Stem Cells through Nuclear Receptor Signaling.- Obtaining Crystals of PPARGamma Ligand Binding Domain Bound to Small Molecules.- Molecular Modeling Approach to Study the PPARGamma-Ligand Interactions.- High Dimensional Data Approaches to Understanding Nuclear Hormone Receptor Signaling.
Nuclear Receptors: A Historical Perspective.- An Optimized Immunoblotting Protocol for Accurate Detection of Endogenous PGC-1 Isoforms in Various Rodent Tissues.- Sub-Cellular Localization of NR4A2 Orphan Nuclear Receptor Expression in Human and Mouse Synovial Joint Tissue.- In Vivo Quantitation of Estrogen Receptor b Subtype Expression in Ovarian Surface Epithelium Using Immunofluorescence Profiling and Confocal Microscopy.- In VivoChIP-Seq of Nuclear Receptors: A Rough Guide to Transform Frozen Tissues into High Confidence Genome-Wide Binding Profiles.- High-Content Analysis of Constitutive Androstane Receptor Nuclear Translocation.- Antibody Validation Strategy for Nuclear Receptors.- Immunofluorescence Labeling of Nuclear Receptor Expression in Formalin-Fixed Paraffin-Embedded Tissue.- Detection of ADP-Ribosylation of the Androgen Receptor Using the Recombinant Macrodomain AF1521 from Archaeoglobus fulgidus.- Reconstitution of the Steroid ReceptorHeterocomplex.- High-Throughput Imaging of PPIX Using Confocal Microscopy.- PGC-1 Overexpression via Local In Vivo Transfection in Mouse Skeletal Muscle.- A Reverse Transfection Method for Screening of Nuclear Receptor Activators.- Hybrid Reporter Gene Assays: Versatile In Vitro Tools to Characterize Nuclear Receptor Modulators.- Analysis of the Transcriptional Activity of Retinoic Acid-Related Orphan Receptors (RORs) and Inhibition by Inverse Agonists.- Examining the Role of Nuclear Receptors during In Vivo Chemical-Mediated Breast Tumorigenesis.- Analysis of IL-4/STAT6 Signaling in Macrophages.- Isolation and Characterization of Adipose Tissue Macrophages.- Assessing Mitochondrial Bioenergetics in Isolated Mitochondria from Mouse Heart Tissues Using Oroboros 2k-Oxygraph.- Generation of Skeletal Myocytes from Embryonic Stem Cells through Nuclear Receptor Signaling.- Obtaining Crystals of PPARGamma Ligand Binding Domain Bound to Small Molecules.- Molecular Modeling Approach to Study the PPARGamma-Ligand Interactions.- High Dimensional Data Approaches to Understanding Nuclear Hormone Receptor Signaling.
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