This book covers techniques in cellular adhesion. It features lists of the necessary materials and reagents; step-by-step, readily reproducible protocols; and notes on troubleshooting and avoiding known pitfalls.

Cellular adhesion is a fundamental process that influences numerous biological activities such as morphogenesis, cell motility and division, as well as signalling. In addition, adhesion is a process important not only in normal physiology and development, but also in disease states such as tumourigenesis, cardiovascular disease, inflammation and infection. There are a plethora of proteins involved in adhesion-related events with a huge diversity in function. As a result, a wide variety of techniques exist to study adhesion related proteins and processes. In Adhesion Protein Protocols, Third Edition, chapters cover techniques to gain insight into the complex and incompletely understood processes that are involved in cellular adhesion. 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.

Authoritative and easily accessible, Adhesion Protein Protocols, Third Edition will be useful for both those new to the field of adhesion protein research as well as the more experienced scientist.

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

Reconstructing Integrin Activation in vitro.- Quantifying Cellular Adhesion to Covalently Immobilized Extracellular Matrix Proteins by Single-Force Spectroscopy.- A Multimode-TIRFM & Microfluidic Technique to Examine Platelet Adhesion Dynamics.- Localization-Based Super-Resolution Imaging of Cellular Structures.- Use of Microarray Analysis to Investigate EMT Gene Signatures.- Podosome Reformation in Macrophages: Assays and Analysis.- Mobile and Three-Dimensional Presentation of Adhesion Proteins Within Microwells.- Basement Membrane Invasion Assays: Native Basement Membrane and Chemoinvasion Assay.- Cytoplasmic Actin: Purification and Single Molecule Assembly Assays.- Synthetic and Tissue-Derived Models for Studying Rigidity Effects on Invadopodia Activity.- Determining Rho GTPase Activity by an Affinity-Precipitation Assay.- Proteomic and Biochemical Methods to Study the Cytoskeletome. Fabricating Surfaces with Distinct Geometries and Different Combinations of Cell Adhesion Proteins.- Purification of Native Arp2/3 Complex from Bovine Thymus.- Purification of Arp2/3 Complex from Saccharomyces Cerevisiae.- Measurement and Analysis of in vitro Actin Polymerization.- Use of the xCELLigence System for Real Time Analysis of changes in Cellular Motility and Adhesion in Physiological Conditions.- Measuring Chemotaxis Using Direct Visualisation Microscope Chambers.- In Vitro Microtubule Severing Assays.- VE-Cadherin Status asan Indicator of Microvascular Permeability.- High-Resolution Live-Cell Imaging and Time-Lapse Microscopy of Invadopodium Dynamics and Tracking Analysis.- Live Cell Imaging of RhoGTPase Biosensors in Tumor Cells.- Electrospun Nanofiber Scaffolds for Investigating Cell-matrix Adhesion.- Method for Measuring Single-Molecule Adhesion Forces and Attachment Lifetimes of Protein-Membrane Interactions.- VE-Cadherin Status as an Indicator of Microvascular Permeability.- High-Resolution Live-Cell Imaging and Time-Lapse Microscopy of Invadopodium Dynamics and Tracking Analysis. Live Cell Imaging of RhoGTPase Biosensors in Tumor Cells.- Electrospun Nanofiber Scaffolds for Investigating Cell-matrix Adhesion.- Method for Measuring Single-Molecule Adhesion Forces and Attachment Lifetimes of Protein-Membrane Interactions.