Adhesion Protein Protocols

1. Front Matter

   Pages i-xi

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2. Monoclonal Antibodies Specific for Leukocyte Adhesion Molecules

   • Carlos Cabañas, Francisco Sánchez-Madrid

   Pages 1-9

3. Epitope Mapping

   • Horace M. DeLisser

   Pages 11-20

4. Sequencing of Antibodies

   • Annie Jarrin, Annie Andrieux

   Pages 21-28

5. Purification Through Affinity Chromatography and Microsequencing

   • Cecilia Garlanda

   Pages 29-37

6. Functional Analysis of Cell Adhesion Molecules

   • David L. Simmons

   Pages 39-63

7. Application of Polymerase Chain Reaction for the Discovery of New Adhesion Molecule Family Members

   • Amy S. Woodard, Lucia R. Languino

   Pages 65-76

8. Construction and Purification of Adhesion Molecule Immunoglobulin Chimeric Proteins

   • Thomas Moll, Dietmar Vestweber

   Pages 77-84

9. Immunofluorescence of Cultured Cells

   • Pier Carlo Marchisio, Livio Trusolino

   Pages 85-92

10. Detection of Adhesion Molecules by Immunohistochemistry on Human and Murine Tissue Sections

    • Antonella Stoppacciaro, Luigi P. Ruco

    Pages 93-106

11. In situ Hybridization with RNA Probes

    • Georg Breier

    Pages 107-117

12. Purification of Fibronectin from Human Plasma

    • Saverio Francesco Retta, Piercarlo Ferraris, Guido Tarone

    Pages 119-124

13. Adhesion to Matrix Proteins

    • Saverio Francesco Retta, Massimo Ternullo, Guido Tarone

    Pages 125-130

14. Leukocyte-Endothelial Monolayer Adhesion Assay (Static Conditions)

    • Jeanne-Marie Kiely, Francis W. Luscinskas, Michael A. Gimbrone Jr.

    Pages 131-136

15. Cell-Cell Adhesive Interactions in an In Vitro Flow Chamber

    • Douglas J. Goetz, Daniel M. Greif, Jian Shen, Francis W. Luscinskas

    Pages 137-145

16. Adhesion of Tumor Cells to Endothelium Under Static Conditions

    • Renato G. S. Chiarivi, Raffaella Giavazzi

    Pages 147-151

17. Adhesion of Tumor Cells Under Flow

    • Andrea Remuzzi, Raffaella Giavazzi

    Pages 153-157

18. Platelet Adhesion to the Subendothelium Under Flow

    • Philip G. de Groot, Martin J. W. IJsseldijk, Jan J. Sixma

    Pages 159-170

19. Leukocyte Transmigration Through Vascular Endothelium

    • Paola Allavena, Aldo Del Maschio

    Pages 171-176

20. Cell Migration into a Wounded Area In Vitro

    • Maria Grazia Lampugnani

    Pages 177-182

Adhesion molecules are of fundamental importance in the regulation of immunity, inflammation, tissue remodeling, and embryonic development. They comprise different families of homologous proteins, such as selectins, integrins, cadherins, and immunoglobins. In addition, beyond these groups, other str- tures with adhesive properties, such as proteoglycans, occludin, and CD44, have been characterized recently. An understanding of the type and characteristics of adhesive molecules expressed by the different cell types and the possibility of manipulating their activity promises considerable clinical potential. Antibodies, small peptidic and nonpeptidic molecules, have recently been used to inhibit thrombosis by blocking platelet aggregation or inflammation through inhibition of leukocyte infiltration and adhesion. Inhibitors of adhesive molecules are used in expe- mental systems for the study of tumor growth and dissemination. Among major goals in the field are the identification of new members of the known adhesive protein families and of independent new adhesive structures. After structural characterization, even more demanding is the study of the biological activity of the new proteins, and the development of simple, rapid tests for the screening of possible inhibitors. In this regard, the production of such reagents as fragments and antibodies would help define the structure–function relati- ship of individual proteins. Data available in the literature show the complexity of the adhesive process and how different molecular epitopes might contribute to the adhesive properties of a single structure. Finally, a new area of investi- tion is the characterization of the intracellular signaling cascade triggered by the engagement of transmembrane adhesive proteins.

"I think this book presents an excellent manual not only for beginning graduate students, who can find a variety of basic techniques, but also for experienced researchers who are new in the field of endothelial biology. . .a very useful manual, containing almost every technique dealing with the cell adhesion properties of the endothelium."-Doody's Health Sciences Book Review Journal

"Dejana and Corada have assembled a wide range of chapters covering methods used for the study of adhesion proteins. The individual chapters are generally well written, and most of the methods that would be expected in such a volume are covered."-Cell Biology International

• Vascular Biology Laboratory, Istituto di Richerche Farmacologie “Mario Negri”, Milano, Italy

  Elisabetta Dejana, Monica Corada

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