XVI, 529 p. 97 illus., 10 illus. in color. online resource
SUMMARY
Integrins play pivotal roles not only across a wide range of physiological processes including tissue morphogenesis, immune responses, wound healing, and regulation of cell growth and differentiation, but also in numerous pathological phenomena such as autoimmunity, thrombosis, and cancer metastasis/progression. Therefore, investigations on integrins often demand multi-disciplinary approaches, making researchers long for a handy collection of comprehensive and practical protocols that detail experimental methods for studying integrin and related cell adhesion molecule functionality. Integrin and Cell Adhesion Molecules: Methods and Protocols aims to provide readers not only with basic protocols in studying integrin functions, but also with summaries on those state-of-the-art technologies that have been utilized for understanding integrin functionality at the cellular, molecular, structural, and organismal levels. Divided into six convenient sections, this detailed volume covers basic protocols for the study of integrin and related cell adhesion molecule functionality in vitro, illustrates structural biology approaches for studying integrins and related cell adhesion molecules, focuses on emerging imaging technologies for investigating cell migration, presents strategies to elucidate signaling through cell adhesion molecules, includes experimental techniques to investigate integrin functions at organismal levels in a physiological context, and showcases the most promising methods and technologies for the development of novel therapeutics and diagnostics. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Both experts and non-experts in the scientific community who wish to study cell adhesion molecules and diagnostics will find Integrin and Cell Adhesion Molecules: Methods and Protocols authoritative, easily accessible, and vastly informative
CONTENT
Overview: Assays for Studying Integrin-Dependent Cell Adhesion -- Cell Adhesion Assays -- Real Time Analysis of Integrin-dependent Transendothelial Migration and Integrin-independent Interstitial Motility of Leukocytes -- Lentiviral Gene Transfer Method to Study Integrin Function in T Lymphocytes -- Surface Plasmon Resonance Biosensing in Studies of the Binding between b2 Integrin I Domains and their Ligands -- Cell-Free Ligand-Binding Assays for Integrin LFA-1 -- Overview: Structural Biology of Integrins -- Protein Expression and Purification of Integrin I Domains and IgSF Ligands for Crystallography -- Electron Microscopic Imaging of Integrin -- An NMR Method to Study Protein-Protein Interactions -- Single Molecule Methods to Study Cell Adhesion Molecules -- Overview: Imaging in the Study of Integrins -- Live Imaging of LFA-1-dependent T Cell Motility and Stop Signals -- Monitoring Integrin Activation by Fluorescence Resonance Energy Transfer -- High-resolution Fluorescence Microscopy to Study Transendothelial Migration -- Multiphoton Intravital Microscopy to Study Lymphocyte Motility in Lymph Nodes -- Overview of Integrin Signaling in the Immune System -- Rap1 and Integrin Inside-out Signaling -- Isolation of Focal Adhesion Proteins for Biochemical and Proteomic Analysis -- Talin and Signaling Through Integrins -- Proteomics Method for Identification of Pseudopodium Phosphotyrosine Proteins -- Overview: Studying Integrins In Vivo -- A Method for the Generation of Conditional Gene-Targeted Mice -- T Cell Homing to the Gut Mucosa: General Concepts and Methodological Considerations -- In Vivo Quantitative Proteomics: the SILAC-Mouse -- Analysis of Chemotaxis in Dictyostelium -- Integrins in Tumor Angiogenesis & Lymphangiogenesis -- PET-Radioimmunodetection of Integrins: Imaging Acute Colitis Using a 64Cu-Labeled Anti-b7 Integrin Antibody -- Integrin-targeted Nanoparticles for siRNA Delivery.- Humanized Mice for Studying Human Leukocyte Integrins In Vivo
