Ion Channels

Methods and Protocols

Editors:

ISBN: 978-1-62703-350-3 (Print) 978-1-62703-351-0 (Online)
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Table of contents (32 protocols)

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  1. Front Matter

    Pages i-xiv

  2. Cloning and Heterologous Expression of Ion Channel Genes

    1. Front Matter

      Pages 1-1

    2. Book Protocol

      Pages 3-19

      Approaches to Cloning of Pain-Related Ion Channel Genes

    3. Book Protocol

      Pages 21-32

      Mammalian Expression Systems and Transfection Techniques

    4. Book Protocol

      Pages 33-54

      Use of Escherichia coli for the Production and Purification of Membrane Proteins

    5. Book Protocol

      Pages 55-64

      Transient Overexpression of Genes in Neurons Using Nucleofection

    6. Book Protocol

      Pages 65-75

      Viral Gene Delivery: Optimized Protocol for Production of High Titer Lentiviral Vectors

  3. Electrophysiological Methods to Study Ion Channel Functions

    1. Front Matter

      Pages 77-77

    2. Book Protocol

      Pages 79-89

      Two-Electrode Voltage Clamp

    3. Book Protocol

      Pages 91-107

      Conventional Micropipette-Based Patch Clamp Techniques

    4. Book Protocol

      Pages 109-118

      Recording of Ion Channel Activity in Planar Lipid Bilayer Experiments

    5. Book Protocol

      Pages 119-131

      Recording Macroscopic Currents in Large Patches from Xenopus Oocytes

    6. Book Protocol

      Pages 133-147

      Combined Single-Channel and Macroscopic Recording Techniques to Analyze Gating Mechanisms of the Large Conductance Ca2+ and Voltage Activated (BK) Potassium Channel

    7. Book Protocol

      Pages 149-157

      Perforated Whole-Cell Patch-Clamp Recording

    8. Book Protocol

      Pages 159-170

      Piezo-Electrically Driven Mechanical Stimulation of Sensory Neurons

    9. Book Protocol

      Pages 171-187

      Automated Planar Patch-Clamp

    10. Book Protocol

      Pages 189-197

      Recording Single-Channel Currents Using “Smart Patch-Clamp” Technique

  4. Imaging and Fluorescence Methods to Study Ion Channels

    1. Front Matter

      Pages 199-199

    2. Book Protocol

      Pages 201-208

      Using Total Internal Reflection Fluorescence Microscopy to Observe Ion Channel Trafficking and Assembly

    3. Book Protocol

      Pages 209-216

      Förster Resonance Energy Transfer-Based Imaging at the Cell Surface of Live Cells

    4. Book Protocol

      Pages 217-231

      The Use of Dansyl-Calmodulin to Study Interactions with Channels and Other Proteins

    5. Book Protocol

      Pages 233-241

      Imaging and Quantification of Recycled KATP Channels

  5. Biochemical and Structure-Functional Approaches in Ion Channel Studies

    1. Front Matter

      Pages 243-243

    2. Book Protocol

      Pages 245-256

      Generation of Antibodies That Are Externally Acting Isoform-Specific Inhibitors of Ion Channels

    3. Book Protocol

      Pages 257-266

      Site-Directed Mutagenesis to Study the Structure–Function Relationships of Ion Channels

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