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This detailed volume covers diverse aspects of P2X7 receptor analysis, ranging from its molecular structure to related pharmacological and immunological tools, via its analysis in heterologous expression systems as well as assays using primary cells and whole animal models. After three introductory chapters that focus on its structure, ligands, and physiological functions, the book details the generation of antibody and nanobody tools for P2X7 receptors, provides protocols for the analysis of expressed P2X7 receptors with a focus on their electrophysiological analysis, as well as protocols for the investigation of P2X7 down-stream signaling in immune cells by flow cytometry. Mouse models and procedures suited to investigate P2X7-mediated effects in other primary cells and in vivo are also explained. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, The P2X7 Receptor: Methods and Protocols is a valuable reference not only for the growing community fascinated by this unusual ion channel but also for a broad readership interested in ion channels or purinergic receptors.
How Structural Biology Has Directly Impacted Our Understanding of P2X Receptor Function and Gating
Agonists, Antagonists, and Modulators of P2X7 Receptors
Modulation of Cell Energy Metabolism by the P2X7 Receptor
Fully Flexible Ligand Docking for the P2X7 Receptor Using ROSIE
Preparation of the Murine Anti-Human P2X7 Receptor Monoclonal Antibody (Clone L4)
Development of Antibody and Nanobody Tools for P2X7
Enhanced Transduction of P2X7-Expressing Cells with Recombinant rAAV Vectors
Immunofluorescence Staining of P2X7 Receptors in Whole-Mount Myenteric Plexus Preparations
Established Protocols for cRNA Expression and Voltage-Clamp Characterization of the P2X7 Receptor in Xenopus laevis Oocytes
A Simplified Protocol to Incorporate the Fluorescent Unnatural Amino Acid ANAP into Xenopus laevis Oocyte-Expressed P2X7 Receptors
Using Whole-Cell Electrophysiology and Patch-Clamp Photometry to Characterize P2X7 Receptor Currents
Using Symmetrical Organic Cation Solutions to Study P2X7 Ion Permeation
Methods for Studying Cholesterol-Dependent Regulation of P2X7 Receptors
Visualizing P2X7-Dependent Inflammasome Formation in Human Monocytes by Fluorescence Microscopy and Flow Cytometry
Characterization of P2X7 Receptors in Human Blood Cells
Real-Time Flow Cytometry as a Tool to Monitor Cellular Consequences of P2X7 Activation in Multiple Cell Populations Mixed in a Single FACS Tube
Administration of P2X7 Receptor Blockers in Oncological Experimental Models
Humanized Mouse Model to Study the P2X7 Receptor in Graft-Versus-Host Disease
Analysis of P2X7-Induced Neuronal Branching
Studying the Role of P2X7 Receptor in Axonal Growth Using In Utero Electroporation Technique
Analyzing the Role of the P2X7 Receptor in Epilepsy