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Immunology has made significant progress in the past decade, driven forward by rapidly advancing technology and a renewed interest in the vast realm of innate immunity. The receptors that mediate these functions are at the front lines of both protective and regulative roles of the immune system. In Immune Receptors: Methods and Protocols, expert researchers present a variety of experimental approaches to the characterization of immune receptors and the cell biology that mediates their functions. These include imaging techniques that aim to understand receptor localization and trafficking, techniques to measure receptor-ligand interactions, strategies to identify novel ligands and methods to analyze downstream receptor signaling, as well as strategies for genomic and proteomic characterization of receptor repertoires. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters include introductions to their respective subjects, 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, Immune Receptors: Methods and Protocols offers technical descriptions and protocols that will be useful both to investigators who are interested in carrying out these procedures and to those who seek a deeper understanding of the bench science that lies behind the immunology literature.
List of contents
Screening for Ligands of C-Type Lectin-Like Receptors.- Yeast Surface Display of Lamprey Variable Lymphocyte Receptors (VLR).- Identification of Scavenger Receptor Ligands.- Construction, Expression, and Purification of Chimeric Protein Reagents Based on Immunoglobulin Fc Regions.- Innate Immune Receptors for Nucleic Acids.- Analysis of Receptor-Ligand Interactions by Surface Plasmon Resonance.- Cell-Based Reporter Assay to Analyze Activation of Nod1 and Nod2.- Determining Fc RI Diffusional Dynamics Via Single Quantum Dot Tracking.- Ratiometric Analysis of Subcellular Recruitment of Fc Receptors During Phagocytosis.- Assessment of the Recycling of the Membrane-Bound Chemokine, CX3CL1.- Measuring Immune Receptor Mobility by Fluorescence Recovery after Photobleaching.- Probing the Plasma Membrane Structure of Immune Cells through the Analysis of Membrane Sheets by Electron Microscopy.- Rapamycin-Based Inducible Translocation Systems for Studying Phagocytosis.- Micropatterned Ligand Arrays to Study Spatial Regulation in Fc Receptor Signaling.- CELLISA: Reporter Cell-Based Immunization and Screening of Hybridomas Specific for Cell Surface Antigens.- Use of Colloidal Silica-Beads for the Isolation of Cell-Surface Proteins for Mass Spectrometry-Based Proteomics.- Transfection-Based Genomic Readout for Identifying Rare Transcriptional Splice Variants.- Characterizing Somatic Hypermutation and Gene Conversion in the Chicken DT40 Cell System.- Characterizing Immune Receptors from New Genome Sequences.
Summary
Immunology has made significant progress in the past decade, driven forward by rapidly advancing technology and a renewed interest in the vast realm of innate immunity. The receptors that mediate these functions are at the front lines of both protective and regulative roles of the immune system. In Immune Receptors: Methods and Protocols, expert researchers present a variety of experimental approaches to the characterization of immune receptors and the cell biology that mediates their functions. These include imaging techniques that aim to understand receptor localization and trafficking, techniques to measure receptor-ligand interactions, strategies to identify novel ligands and methods to analyze downstream receptor signaling, as well as strategies for genomic and proteomic characterization of receptor repertoires. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective subjects, 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, Immune Receptors: Methods and Protocols offers technical descriptions and protocols that will be useful both to investigators who are interested in carrying out these procedures and to those who seek a deeper understanding of the bench science that lies behind the immunology literature.
