| C1858 Sigma |
Connexin Methods and Protocols |
General description
Volume 154 in Methods in Molecular Biology Series describes the best techniques to study cellular and molecular biology of connexins, as well as physiological properties. These methods are specifically designed to investigate the life of connexins from biosynthesis to assembly into gap junction channels, and include powerful assays to perform functional studies on these routes of cell-cell communication. It offers a collection of methods for studying the role of connexins in intercellular communication and human genetic disorders, as well as finding new therapeutics.
Table of Contents
PART I. TOOLS TO STUDY CONNEXINS.
Investigation of Connexin Gene Expression Patterns by In Situ Hybridization Techniques,
Sodium Dodecyl Sulfate-Freeze-Fracture Immunolabeling of Gap Junctions,
Purification of Gap Junctions,
Culturing of Mammalian Cells Expressing Recombinant Connexins and Two-Dimensional Crystallization of the Isolated Gap Junctions,
Connexins/Connexons: Cell-Free Expression,
Biochemical Analysis of Connexon Assembly,
Expression and Imaging of Connexin-GFP Chimeras in Live Mammalian Cells,
Analysis of Connexin Expression in Brain Slices by Single-Cell Reverse Transcriptase Polymerase Chain Reaction, .
Use of Retroviruses to Express Connexins,
Spatiotemporal Depletion of Connexins Using Antisense Oligonucleotides,
Transfection and Expression of Exogenous Connexins in Mammalian Cells,
PART II. ASSAYS FOR FUNCTION
Assaying the Molecular Permeability of Connexin Channels,
Applying the Xenopus Oocyte Expression System to the Analysis of Gap Junction Proteins,
Mutagenesis to Study Channel Structure,
Dual Patch Clamp,
Determining Ionic Permeabilities of Gap Junction Channels,
Fluorescence Recovery After Photobleaching,
Capture of Transjunctional Metabolites,
PART III. PHYSIOLOGY AND BIOLOGY OF CONNEXINS
The Study of Connexin Hemichannels (Connexons) in Xenopus Oocytes,
Exploring Hemichannel Permeability In Vitro,
Inducing De Novo Formation of Gap Junction Channels,
Recording and Analysis of Putative Direct Electrical Interactions in the Mammalian Brain,
Intercellular Calcium Signaling and Flash Photolysis of Caged Compounds: A Sensitive Method to Evaluate Gap Junctional Coupling,
Biochemical Analysis of Connexin Phosphorylation,
How to Close a Gap Junction Channel: Efficacies and Potencies of Uncoupling Agents,
Investigation of Connexin Gene Expression Patterns by In Situ Hybridization Techniques,
Sodium Dodecyl Sulfate-Freeze-Fracture Immunolabeling of Gap Junctions,
Purification of Gap Junctions,
Culturing of Mammalian Cells Expressing Recombinant Connexins and Two-Dimensional Crystallization of the Isolated Gap Junctions,
Connexins/Connexons: Cell-Free Expression,
Biochemical Analysis of Connexon Assembly,
Expression and Imaging of Connexin-GFP Chimeras in Live Mammalian Cells,
Analysis of Connexin Expression in Brain Slices by Single-Cell Reverse Transcriptase Polymerase Chain Reaction, .
Use of Retroviruses to Express Connexins,
Spatiotemporal Depletion of Connexins Using Antisense Oligonucleotides,
Transfection and Expression of Exogenous Connexins in Mammalian Cells,
PART II. ASSAYS FOR FUNCTION
Assaying the Molecular Permeability of Connexin Channels,
Applying the Xenopus Oocyte Expression System to the Analysis of Gap Junction Proteins,
Mutagenesis to Study Channel Structure,
Dual Patch Clamp,
Determining Ionic Permeabilities of Gap Junction Channels,
Fluorescence Recovery After Photobleaching,
Capture of Transjunctional Metabolites,
PART III. PHYSIOLOGY AND BIOLOGY OF CONNEXINS
The Study of Connexin Hemichannels (Connexons) in Xenopus Oocytes,
Exploring Hemichannel Permeability In Vitro,
Inducing De Novo Formation of Gap Junction Channels,
Recording and Analysis of Putative Direct Electrical Interactions in the Mammalian Brain,
Intercellular Calcium Signaling and Flash Photolysis of Caged Compounds: A Sensitive Method to Evaluate Gap Junctional Coupling,
Biochemical Analysis of Connexin Phosphorylation,
How to Close a Gap Junction Channel: Efficacies and Potencies of Uncoupling Agents,
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