Adhesion Protein Protocols

The second edition of Adhesion Protein Protocols combines traditional techniques with cutting-edge and novel techniques that can be adapted easily to different molecules and cell types. The topics discussed include novel techniques for studying cell-cell adhesion, neutrophil chemotaxis, in vitro assays used to study leukocyte migration through monolayers of cultured endothelial cells, and novel techniques to purify pseudopodia from migratory cells. The protocols discussed in this volume are suitable for both novice and expert scientists, who will gain further insight into the complex and incompletely understood processes involved in cellular adhesion.

Methods in Molecular Biology Volume 96: Adhesion Protocols

Editor: Dr Amanda S. Coutts

Table of Contents

Preface



1. Manipulation of cell-cell adhesion using bowtie-shaped microwells. Celeste M. Nelson, Wendy F. Liu, and Christopher S. Chen.



2. Separation of cell-cell adhesion complexes by differential centrifugation. Roger Vogelmann and W. James Nelson.



3. Analysis of neutrophil chemotaxis. Paul A. Nuzzi, Mary A. Lokuta, and Anna Huttenlocher.



4. Analysis of leukocyte migration through monolayers of cultured endothelial cells. Helen M. McGettrick, Lynn M. Butler, and Gerard B. Nash.



5. Biochemical purification of pseudopodia from migratory cells. Yingchun Wang and Richard L. Klemke.



6. Dynamic assessment of cell-matrix mechanical interactions in 3-D culture. W. Matthew Petroll.



7. Quantitative Analyses of Cell Adhesion Strength. Nathan D. Gallant and Andrés J. García



8. Using RNA interference to knockdown the adhesion protein TES. Elen Griffith.



9. Assaying calpain activity. Neil O. Carragher.



10. Analysis of focal adhesions and cytoskeleton by custom microarray. Matthew J. Dalby and Stephen J. Yarwood.



11. Proteomic analysis of cell surface membrane proteins in leukemic cells. Robert S. Boyd, Martin J.S. Dyer, and Kelvin Cain.



12. Bioinformatic analysis of adhesion proteins. Josephine C. Adams and Juergen Engel.



13. Analysis of integrin dynamics by Fluorescence Recovery after Photobleaching. Bernard Wehrle-Haller.



14. Double-Hydrogel Substrate as a Model System for 3D Cell Culture. Karen A. Beningo and Yu-li Wang.



15. In vitro actin assembly assays and purification from Acanthamoeba. J. Bradley Zuchero.



Subject Index