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This Methods in Molecular Biology(TM) volume covers basic protocols for the study of integrin and related cell adhesion molecule functionality, and showcases the most promising methods and technologies for the development of novel therapeutics and diagnostics.
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.
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Inhalt
Part I: Basic Protocols for the Study of Integrin and Related Cell Adhesion Molecule Functionality In Vitro
Overview: Assays for Studying Integrin-Dependent Cell Adhesion
Alexandre Chigaev and Larry A. Sklar
Cell Adhesion Assays
Gabriele Weitz-Schmidt and Stéphanie Chreng
Real Time Analysis of Integrin-dependent Transendothelial Migration and Integrin-independent Interstitial Motility of Leukocytes
Ziv Shulman and Ronen Alon
Lentiviral Gene Transfer Method to Study Integrin Function in T Lymphocytes
Daliya Banerjee and Motomu Shimaoka
Surface Plasmon Resonance Biosensing in Studies of the Binding between b2 Integrin I Domains and their Ligands
Thomas Vorup-Jensen
Cell-Free Ligand-Binding Assays for Integrin LFA-1
Koichi Yuki
Part II: Structural Biology Approaches for Studying Integrins and Related Cell Adhesion Molecules
Overview: Structural Biology of Integrins
Guanyuan Fu, Wei Wang, and Bing-Hao Luo
Protein Expression and Purification of Integrin I Domains and IgSF Ligands for Crystallography
Hongmin Zhang and Jia-huai Wang
Electron Microscopic Imaging of Integrin
Kenji Iwasaki
An NMR Method to Study Protein-Protein Interactions
Noritaka Nishida and Ichio Shimada
Single Molecule Methods to Study Cell Adhesion Molecules
Joonil Seog
Part III: Imaging for Investigating Cell Adhesion and Migration
Overview: Imaging in the Study of Integrins
Christopher V. Carman
Live Imaging of LFA-1-dependent T Cell Motility and Stop Signals
Andrew J. Wiemer, Sarah Wernimont, and Anna Huttenlocher
Monitoring Integrin Activation by Fluorescence Resonance Energy Transfer
Craig T. Lefort, Young-min Hyun, and Minsoo Kim
High-resolution Fluorescence Microscopy to Study Transendothelial Migration
Christopher V. Carman
Multiphoton Intravital Microscopy to Study Lymphocyte Motility in Lymph Nodes
Thomas T. Murooka and Thorsten R. Mempel
Part IV: Signaling through Cell Adhesion Molecules
Overview of Integrin Signaling in the Immune System
Tatsuo Kinashi
Rap1 and Integrin Inside-out Signaling
Koko Katagiri and Tatsuo Kinashi
Isolation of Focal Adhesion Proteins for Biochemical and Proteomic Analysis
Jean-Cheng Kuo, Xuemei Han, John R. Yates III, and Clare M. Waterman
Talin and Signaling Through Integrins
Mohamed Bouaouina, David S. Harburger, and David A. Calderwood
Proteomics Method for Identification of Pseudopodium Phosphotyrosine Proteins
Yingchun Wang and Richard L. Klemke
Part V: Cell Adhesion and Migration at Organismal Levels in a Physiological Context
Overview: Studying Integrins In Vivo
Clifford A. Lowell and Tanya N. Mayadas
A Method for the Generation of Conditional Gene-Targeted Mice
Masahiro Yamamoto and Kiyoshi Takeda
T Cell Homing to the Gut Mucosa: General Concepts and Methodological Considerations
Jaime De Calisto, Eduardo J. Villablanca, Sen Wang, Maria R. Bono, Mario Rosemblatt, and J. Rodrigo Mora
In Vivo Quantitative Proteomics: the SILAC-Mouse
Sara Zanivan, Marcus Krueger, and Matthias Mann
Analysis of Chemotaxis in Dictyostelium
Huaqing Cai, Chuan-Hsiang Huang, Peter N. Devreotes, and Miho Iijima
Part VI: Methods and Technologies towards Novel Therapeutics and Diagnostics Targeting Integrins
Integrins in Tumor Angiogenesis & Lymphangiogenesis
Philippe Foubert and Judith A. Varner
PET-Radioimmunodetection of Integrins: Imaging Acute Colitis Using a 64Cu-Labeled Anti-b7 Integrin Antibody
Jason L.J. Dearling and Alan B. Packard
Integrin-targeted Nanoparticles for siRNA Delivery
Noa Ben-Arie, Ranit Kedmi, and Dan Peer
Humanized Mice for Studying Human Leukocyte Integrins In Vivo
Sang-Soo Kim, Priti Kumar, and Premlata Shankar