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.
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.
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