Integrin Protocols

ISBN-13: 9781489943415 / Angielski / Miękka / 2013 / 296 str.

Anthony R. Howlett

Opis

It is now widely accepted that the extracellular matrix (ECM) is a key determinant of tissue-specific gene expression. Signals provided by ECM are transduced by integrins, a large and growing superfamily of transmembrane heterodimeric cell surface receptors that link the ECM to structural and fu- tional elements within the cell. A wide range of cellular phenotypes have been shown to be regulated by integrins, including growth, differentiation, mig- tion, invasion, angiogenesis, and apoptosis. Furthermore, abnormalities of integrin expression and function have been implicated in the etiology of va- ous pathologic conditions, including cardiovascular disease, inflammatory disorders, and cancer. Thus integrins have emerged as an important class of molecules with wide ranging implications for understanding basic biological processes. In Integrin Protocols we provide a wide-ranging collection of laboratory protocols intended to assist investigators interested in integrins in working productively with these molecules, in studying their expression, and in pot- tially manipulating that expression to define their role(s) in relevant biolo- cal models. Protocols are provided for the analysis of integrin expression both at the RNA and protein levels (Chaps. 2, 5, and 7). Delcommenne and Streuli describe procedures for making rat monoclonal antibodies specific for mouse integrins; Schneller et al. and Arap and Huang describe methods for western blotting of integrins and RT-PCR analysis. Protocols are included that cover the analysis of the functional properties of integrins (Chaps. 1, 3, 4, 8, and 9 through 11). Koivunen et al.

Recenzje

"Integrin Protocols aims to provide a hand-book of methods for the study of integrins. Many of the essential techniques are covered, with sections ranging from the relatively straight-forward (immunoblotting) to the highly complex (phage display and knock-outs)...Over half of the sections are by major figures in the integrin field, thus giving the book considerable authority..."-Cell Biology International 2000

Spis treści

Part 1 Working with Integrins. Integrin-Binding Peptides Derived from Phage Display Libraries, Erkki Koivunen, Bradley H. Restel, Daniel Rajotte, Johanna Lahdenranta, Martin Hagedorn, Wadih Arap, and Renata Pasqualini. Production of Rat Monoclonal Antibodies Specific for Mouse Integrins, Marc Delcommenne and Charles H. Streuli. Biochemical Analysis of Integrin-Mediated Shc Signaling, Kishore K. Wary, Michael Dans, Agnese Mariotti, and Filippo G. Giancotti. Assays for Kinase Activity, Maja H. Oktay, Agnese Mariotti, and Filippo G. Giancotti. Immunoblotting of Integrins, Maximilian Schneller, Wadih Arap, and Renata Pasqualini. Application of Flow Cytometry in the Analysis and Sterile Sorting of Cell Populations Based on Integrin Expression, Edward C. Rosfjord and Robert B. Dickson. Expression of Integrin Transcripts in Human Cancer Cells, Wadih Arap and H.-J. Su Huang. Part 2 Functional Analysis of Integrins. Functional Analysis of the b2 Integrins, Rui Li and M. Amin Arnaout. Expression of Heterologous Integrin Genes, Craig A. Steger, Mara Fornaro, Amy S. Woodard, Duo-Qi Zheng, and Lucia R. Languino. Preparing a Polyclonal Antibody to Mouse b1 Integrin with Function-Blocking Activity, Gwynneth M. Edwards and Charles H. Streuli. Integrin Gene Targeting, Jan F. Talts, Cord Brakebusch, and Reinhard Fässler. Determination of Positive and Negative Regulatory Elements, Emilio Hirsch, Piero Cervella, Lorenzo Silengo, and Fiorella Altruda. Part 3 Integrin Function at the Cellular Level. A Cell-Based Adhesion Assay for the Characterization of Integrin avb3 Antagonists, James M. Wu, Mary P. Rosser, Anthony R. Howlett, and Richard I. Feldman. Integrins in Cell Migration, Klaus von der Mark, Stephan Schöber, and Simon L. Goodman. Models for Studying Cellular Invasion of Basement Membranes, John T. Price and Erik W. Thompson. Methods for Studying Anoikis, Steven M. Frisch. Use of the 10-Day-Old Chick Embryo Model for Studying Angiogenesis, Peter C. Brooks, Anthony M. P.Montgomery, and David A. Cheresh. Cell Shape and Integrin Signaling Regulate the Differentiation State of Mammary Epithelial Cells, Aruna Somasiri and Calvin D. Roskelley. Index.

Blurb

In Integrin Protocols, 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. The emphasis is always on the practical steps necessary for experimental success and robust results.
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.