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Presentingstate-of-the-art protocols to study gap junctions, this detailed book firstfocuses on the use of methods and tools to investigate the different aspects ofconnexin expression and gap junction regulation. The second part of the volumedescribes several methods to probe gap junction functionality as such. Writtenin the highly successful Methods in Molecular Biology series format, chaptersinclude introductions to their respective topics, lists of the necessarymaterials and reagents, step-by-step and readily reproducible laboratoryprotocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritativeand practical, Gap Junction Protocols is intended for basic and appliedresearchers in the area of biomedical and life sciences, both in academic andindustrial settings.
Inhaltsverzeichnis
Analysis of Liver Connexin ExpressionUsing Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction.- DNA Methylation Analysis of HumanTissue-Specific Connexin Genes.- Detection of Connexins in Liver Cells UsingSodiumdodecylsulfate Polyacrylamide Gel Electrophoresis and Immunoblot Analysis.- Immunohisto- andCytochemistry Analysis of Connexins.- Small Interfering RNA-MediatedConnexin Gene Knockdown in Vascular Endothelial and Smooth Muscle Cells.- Generation and Use of TrophoblastStem Cells and Uterine Myocytes to Study the Role of Connexins for Pregnancyand Labor.- Identification of Connexin43Phosphorylation and S-Nitrosylationin Cultured Primary Vascular Cells.- Preparation of Gap Junctions inMembrane Microdomains for Immunoprecipitation and Mass Spectrometry InteractomeAnalysis.- Scrape Loading/Dye Transfer Assay.- Microinjection Technique for Assessment of GapJunction Function.- Electroporation Loading and DyeTransfer: A Safe and Robust Method to Probe Gap Junctional Coupling.- Using Fluorescence Recovery After Photobleaching to Study Gap JunctionalCommunication In Vitro.- Tracking Dynamic Gap Junctional Coupling in Live Cells by LocalPhoto-Activation and Fluorescence Imaging.- A Functional Assay to Assess Connexin43-Mediated Cell-to-Cell Communication ofSecond Messengers in Cultured Bone Cells.- Calcium Wave Propagation Triggered by Local Mechanical Stimulation as a Methodfor Studying Gap Junctions and Hemichannels.- Establishment of the Dual Whole Cell Recording Patch Clamp Configuration forthe Measurement of Gap Junction Conductance.
Zusammenfassung
Presenting
state-of-the-art protocols to study gap junctions, this detailed book first
focuses on the use of methods and tools to investigate the different aspects of
connexin expression and gap junction regulation. The second part of the volume
describes several methods to probe gap junction functionality as such. 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 and readily reproducible laboratory
protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative
and practical, Gap Junction Protocols is intended for basic and applied
researchers in the area of biomedical and life sciences, both in academic and
industrial settings.
