Integrin Protocols

About this book

Anthony Howlett and a distinguished panel of experimentalists describe in detail a series of cutting-edge methods for dissecting the role of integrins in biological processes. This wide-ranging collection includes protocols for the analysis of integrin expression - at both the RNA and protein levels - and for elucidating the functional properties of integrins, including those at the cellular level. Each method provides step-by-step instructions for easy reproducibility, along with extensive notes about potential pitfalls, and tips on how to avoid failure. Offering powerful tools for understanding how integrins regulate cell growth, differentiation, migration, invasion, angiogenesis, and apoptosis as well as how abnormalities of integrin expression and function may be implicated in various pathologic conditions, Integrin Protocols constitutes a gold-standard collection of techniques for both new and experienced investigators of the molecular and cellular basis of cardiovascular disease, inflammatory disorders, and cancer.

Contents

Part I Working with Integrins; Integrin-Binding Pepetides Derived From Phage Display Libraries; Production of Rat Monoclonal Antibodies Specific for Integrins; Biochemical Analysis of Integrin Mediated Shc Signaling. Assays for Kinase Activity; Immunoblotting of Integrins; Application of Flow Cytometry in the Analysis and Sterile Sorting of Cell Populations Based on Integrin Expression; Expression of Integrin Transcripts in Human Cancer Cells; Part II Functional Analysis of Integrins. Functional Analysis of the b2 Integrins; Expression of Heterologous Integrin Genes; Preparing a Polyclonal Antibody to Mouse b1 Integrin with Function-Blocking Activity; Integrin Gene Targeting. Determination of Positive and Negative Regulatory Elements; Part III Integrin Function at the Cellular Level; A Cell-Based Adhesion Assay for the Characterization of Integrin avb3 Antagonists; Integrins in Cell Migration. Models for Studying Cellular Invasion of Basement Membranes; Methods for Studying Anoikis; Use of the Ten-Day-Old Chick Embryo Model for Studying Angiogenesis; Cell Shape and Integrin Signaling Regulate the Differentiation State of Mammary Epithelial Cells

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