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Full Description
This second edition volume expands on the previous edition with new discussions on the latest techniques used to study molecular chaperones and the stress response. The chapters in this book cover such as analysis of the initiation and regulation of the stress response; the role of heat shock protein 90 (Hsp90) in gene expression through chromosome-immunoprecipitation; features of chaperone function and biology; the emerging role of the extracellular HSPs; and the use of chaperones as biomarkers. 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.
Cutting-edge and thorough, Chaperones: Methods and Protocols, Second Edition is a valuable resource for all researchers who want to learn more about this interesting and developing field.
Contents
Monitoring of the Heat Shock Response with a Real-Time Luciferase Reporter.- Studying RNA Polymerase II Promoter-Proximal Pausing by In Vitro Immobilized Template and Transcription Assays.- Role of Heat Shock Factors in Stress-Induced Transcription: An Update.- A Workflow Guide to RNA-seq Analysis of Chaperone Function and Beyond.- Chromatin Immunoprecipitation (ChIP) of Heat Shock Protein 90 (Hsp90).- Transfection and Thermotolerance Methods for Analysis of miR-570 Targeting the HSP Chaperone Network.- Targeted Replacement of HSF1 Phosphorylation Sites at S303/S307 with Alanine Residues in Mice Increases Cell Proliferation and Drug Resistance.- Bimolecular Fluorescence Complementation Assay to Evaluate HSP90-Client Protein Interactions in Cells.- Complementation Assays for Co-Chaperone Function.- Optimized Microscale Protein Aggregation Suppression Assay: A Method for Evaluating the Holdase Activity of Chaperones.- Detecting Post-Translational Modifications of Hsp90 Isoforms.- Multiple Targeting of HSP Isoforms to Challenge Isoform Specificity and Compensatory Expression.- Using a Modified Proximity Ligation Protocol to Study the Interaction between Chaperones and Associated Proteins.- Use of Native-PAGE for the Identification of Epichaperomes in Cell Lines.- Molecular Chaperone Receptors: An Update.- A Novel Heat Shock Protein 70-Based Vaccine Prepared from DC-Tumor Fusion Cells: An Update.- Methods to Assess the Impact of Hsp90 Chaperone Function on Extracellular Client MMP2 Activity.- Proteome and Chaperone Analysis of Extracellular Vesicles.- A Modified Differential Centrifugation Protocol for Isolation and Quantitation of Extracellular Heat Shock Protein 90 (eHsp90).- Immunohistochemistry of Human Hsp60 in Health and Disease: Recent Advances in Immuno-Morphology and Methods for Assessing the Chaperonin in Extracellular Vesicles.- Multiplex Immunostaining Method to Distinguish HSP Isoforms in Cancer Tissue Specimens.- Large-Scale Databases and Portals on Cancer Genome to Analyze Chaperone Genes Correlated to Patient Prognosis.- Immunohistochemical, Flow Cytometric and ELISA-Based Analyses of Intracellular, Membrane-Expressed, and Extracellular Hsp70 as Cancer Biomarkers.
