This volume provides readers with the latest techniques and approaches used to study K+ channels. There are a wide range of topics covered in the book, such as channel structure; K+ channels and their roles in physiology and disease; single-molecule FRET; proteins expression levels in vivo; real-time correlation of channel activity; gating; pharmacology; physiological functions; and channelopathies. 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,…mehr
This volume provides readers with the latest techniques and approaches used to study K+ channels. There are a wide range of topics covered in the book, such as channel structure; K+ channels and their roles in physiology and disease; single-molecule FRET; proteins expression levels in vivo; real-time correlation of channel activity; gating; pharmacology; physiological functions; and channelopathies. 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 comprehensive, Potassium Channels: Methods and Protocols is a valuable resource for all researchers who are interested in studying potassium channels.
Manipulating Potassium Channel Expression and Function in Hippocampal Neurons by In Utero Electroporation.- Studying KCNQ1 Mutation and Drug Response in Type 1 Long QT Syndrome using Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes.- Monitoring Changes in the Abundance of Endogenously Expressed ATP-Sensitive Potassium (KATP) Channels in the Plasma Membrane using Surface Biotinylation.- Nonsense-Mediated mRNA Decay of hERG Mutations in Long QT Syndrome.- Probing Subunits Interactions in KATP Channels using Photo-Crosslinking via Genetically Encoded p-azido-L-phenylalanine.- Hyper-SUMOylation of K+ Channels in Sudden Unexplained Death in Epilepsy: Isolation and Primary Culture of Dissociated Hippocampal Neurons from Newborn Mice for Subcellular Localization.- Simultaneous Real-Time Measurement of the beta-Cell Membrane Potential and Ca2+ Influx to Assess the Role of Potassium Channels on beta-Cell Function.- Methods for Characterizing Disease-Associated ATP-SensitivePotassium Channel Mutations.- Thallium Flux Assay for Measuring the Activity of Monovalent Cation Channels and Transporters.- Nuclear Magnetic Resonance Approaches for Characterizing Protein-Protein Interactions.- Studying Mechanosensitivity of Two-Pore Domain K+ Channels in Cellular and Reconstituted Proteoliposome Membranes.- Migration of PIP2 on KCNQ2 Surface Revealed by Molecular Dynamics Simulations.- Studying Structural Dynamics of the Potassium Channels by Single-Molecule FRET.- Patch-Clamp Recordings of the KcsA K+ Channel in Unilamellar Blisters.- Combinatorial Assembly of Lumitoxins.- Characterization of MC4R Regulation of the Kir7.1 Channel using the Tl+ Flux Assay.- Stopped-Flow Fluorometric Ion Flux Assay for Ligand-Gated Ion Channel Studies.- In Vivo Analysis of Potassium Channelopathies: Loose Patch Recording of Purkinje Cell Firing in Living, Awake Zebrafish.- Site-Directed Unnatural Amino Acid Mutagenesis to Investigate Potassium Channel Pharmacology in Xenopus laevisOocytes.- Random Spherically Constrained Single-Particle (RSC) Method to Study Voltage-Gated Ion Channels.- CW-EPR Spectroscopy and Site-Directed Spin Labeling to Study the Structural Dynamics of Ion Channels.- Ion Binding to Transport Proteins using Isothermal Titration Calorimetry.- Building Atomic Models of the Ion Channels Based on Low Resolution Electron Microscopy Maps and Homology Modeling.- Studying Kv Channels Function using Computational Methods.
Manipulating Potassium Channel Expression and Function in Hippocampal Neurons by In Utero Electroporation.- Studying KCNQ1 Mutation and Drug Response in Type 1 Long QT Syndrome using Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes.- Monitoring Changes in the Abundance of Endogenously Expressed ATP-Sensitive Potassium (KATP) Channels in the Plasma Membrane using Surface Biotinylation.- Nonsense-Mediated mRNA Decay of hERG Mutations in Long QT Syndrome.- Probing Subunits Interactions in KATP Channels using Photo-Crosslinking via Genetically Encoded p-azido-L-phenylalanine.- Hyper-SUMOylation of K+ Channels in Sudden Unexplained Death in Epilepsy: Isolation and Primary Culture of Dissociated Hippocampal Neurons from Newborn Mice for Subcellular Localization.- Simultaneous Real-Time Measurement of the beta-Cell Membrane Potential and Ca2+ Influx to Assess the Role of Potassium Channels on beta-Cell Function.- Methods for Characterizing Disease-Associated ATP-SensitivePotassium Channel Mutations.- Thallium Flux Assay for Measuring the Activity of Monovalent Cation Channels and Transporters.- Nuclear Magnetic Resonance Approaches for Characterizing Protein-Protein Interactions.- Studying Mechanosensitivity of Two-Pore Domain K+ Channels in Cellular and Reconstituted Proteoliposome Membranes.- Migration of PIP2 on KCNQ2 Surface Revealed by Molecular Dynamics Simulations.- Studying Structural Dynamics of the Potassium Channels by Single-Molecule FRET.- Patch-Clamp Recordings of the KcsA K+ Channel in Unilamellar Blisters.- Combinatorial Assembly of Lumitoxins.- Characterization of MC4R Regulation of the Kir7.1 Channel using the Tl+ Flux Assay.- Stopped-Flow Fluorometric Ion Flux Assay for Ligand-Gated Ion Channel Studies.- In Vivo Analysis of Potassium Channelopathies: Loose Patch Recording of Purkinje Cell Firing in Living, Awake Zebrafish.- Site-Directed Unnatural Amino Acid Mutagenesis to Investigate Potassium Channel Pharmacology in Xenopus laevisOocytes.- Random Spherically Constrained Single-Particle (RSC) Method to Study Voltage-Gated Ion Channels.- CW-EPR Spectroscopy and Site-Directed Spin Labeling to Study the Structural Dynamics of Ion Channels.- Ion Binding to Transport Proteins using Isothermal Titration Calorimetry.- Building Atomic Models of the Ion Channels Based on Low Resolution Electron Microscopy Maps and Homology Modeling.- Studying Kv Channels Function using Computational Methods.
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