Adhesion Proteins Protocols

About this book

An international corps of expert investigators describe their optimized techniques for both the identification of new cell adhesion proteins and for the characterization of novel adhesive structures. Their cutting edge methods will enable both novice and experienced researchers readily to identify and clone new adhesion proteins, to successfully produce inhibitors of the adhesive activity, to develop biological models for the assay of cell-to-matrix and cell-to-cell adhesion in vitro, and to measure the signaling activities of adhesion proteins. By illuminating these adhesive molecules and the possibilities for manipulating them, the new experimental strategies collected here will have considerable clinical potential for the regulation of immunity, inflammation, tissue remodeling, and embryonic development.

Contents

Production of Monoclonals Directed to Adhesive Molecules. Monoclonal Antibodies Specific for Leucocyte Adhesion Molecules: Selective Protocols of Immunization and Screening Assays for Generation of Blocking Activating and Activation Reporter Antibodies C. Cabanas and F. Sanchez-Madrid. Epitope Mapping H.M. DeLisser. Sequencing of Antibodies A. Jarrin and A. Andrieux. Isolation and Cloning of Adhesive Molecules. Purification Through Affinity Chromatography and Microsequencing C. Garlanda. Functional Analysis of Cell Adhesion Molecules D. Simmons. Application of Polymerase Chain Reaction for the Discovery of New Adhesion Molecule Family Members A.S. Woodard and L.R. Languino. Construction and Purification of Adhesion Molecule-Immunoglobulin Chimeric Proteins T. Moll and D. Vestweber. Morphological Localization of Adhesive Molecules and Related Cytoskeletal Components. Immunofluorescence of Cultured Cells P.C. Marchisio and L. Trusolino. Detection of Adhesion Molecules by Immunohistochemistry on Human and Murine Tissue Sections A. Stopacciaro and L.P. Ruco. In situ Hybridization with RNA Probes G. Breier. Adhesion of Circulating Cells to the Endothelium and to Extracellular Matrix Proteins. Purification of Fibronectin and Vitronectin from Human Plasma S.F. Retta P. Ferraris and G. Tarone. Adhesion to Matrix Proteins S.F. Retta M. Ternullo and G. Tarone. Leucocyte- Endothelial Monolayer Adhesion Assay (Static Conditions) J.M. Kiely F.W. Luscinskas and M.A. Gimbrone. Cell-Cell Adhesive Interactions in an In Vitro Flow Chamber D.J. Goetz D.M. Greif J. Shen and F.W. Luscinskas. Adhesion of Tumor Cells to Endothelium Under Static Conditions R.G.S. Chiarivi and R. Giavazzi. Adhesion of Tumor Cell Under Flow A. Remuzzi and R. Giavazzi. Platelet Adhesion to the Subendothelium Under Flow. Platelet Adhesion to the Subendothelium Under Flow G. de Groot M.J.W. Ijsseldijk and J.J. Sixma. Cell Chemotaxis. Leucocyte Transmigration Through Vascular Endothelium: An In Vitro Method P. Allavena and A. Del Maschio. Cell Migration into a Wounded Area In Vitro M.G. Lampugnani. Endothelial Cell Permeability In Vitro. Determination of the Endothelial Barrier Function In Vitro G.P. van Nieuw Amerongen and V.W.M. van Hinsbergh. Induction of Intercellular Signals Through Binding of Adhesive Molecules. Activation of p125 FAK Tyrosine Phosphorylation M. Venturino C. Olivo G. Tarone and P. Defilippi. Induction of Ca2+ Flux by Adhesion Molecules in Lymphocytes R. DeMaria G. Palmieri and A. Santoni. Integrin-Mediated Stimulation of Tyrosine Phosphorilation in Lymphoid Cells A. Gismondi and A. Santoni. The Identification of Signaling Molecules by the Yeast 2-Hybrid System S. Biffo and P.C. Marchisio. Index.

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