Leading bench-top experimentalists from around the world present, in a proven format, their state-of-the-art methods for probing the extracellular matrix (ECM), its mechanisms of assembly and how it influences cell behavior. These carefully detailed biochemical, biophysical, genetic, molecular biological, and cell biological techniques have been optimized for the study of ECM structure and assembly, the mechanisms of cell-ECM interactions, and the molecular genetics of ECM proteins and defects in human disease, and bring to a wider audience a range of highly sophisticated methods that have hitherto been difficult even for a few specialized laboratories. Cutting-edge and highly practical, Extracellular Matrix Protocols offers today's ECM researchers a gold-standard collection of powerful, readily reproducible methods to investigate the extracellular matrix and the altered cell-ECM interactions that play so important a role in human health and disease.
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"This would be most useful to a lab researcher who had not previously been studying the role of the ECM in biological systems, and who wished to do so. Having all the practical protocols for preparing, assaying, and studying these important molecules in a small volume could be very useful for such a researcher."-Doody's Health Science Book Review Journal
Part I. Biochemistry of Extracellular Matrix. Semipermeabilized Cells to Study Procollagen Assembly, Richard R. Wilson and Neil J. Bulleid. Quantitative Determination of Collagen Crosslinks, Trevor J. Sims, Nicholas C. Avery, and Allen J. Bailey. Analysis of Laminin Structure and Function with Recombinant Glycoprotein Expressed in Insect Cells, Todd L. Mathus and Peter D. Yurchenco. Recombinant Collagen Trimers from Insect Cells and Yeast, Johanna Myllyharju. Eukaryotic Expression and Purification of Recombinant Extracellular Matrix Proteins Carrying the Strep II Tag, Neil Smyth, Uwe Odenthal, Barbara Merkl, and Mats Paulsson. Part II. Biophysics of Extracellular Matrix Proteins. Preparation of Isotopically Labelled Recombinant Fragments of Fibronectin for Functional and Structural Study by Heteronuclear Nuclear Magnetic Resonance Spectroscopy, Jeremy R. Bright, Andrew R. Pickford, Jennifer R. Potts, and Iain D. Campbell. Reconstitution of Functional Integrin into Phopsholipid Vesicles and Planar Lipid Bilayers, Eva-Maria Erb and Jürgen Engel. Confocal-FRAP Analysis of ECM Molecular Interactions, Timothy Hardingham and Philip Gribbon. Electron Cryomicroscopy of Fibrillar Collagens, Roger S. Meadows, David F. Holmes, Chris J. Gilpin, and Karl E. Kadler. Atomic Force Microspopy Measurements of Intermolecular Binding Strength, Gradimir N. Misevic. ECM Macromolecules: Rotary Shadowing and Scanning Transmission Electron Microscopy, Michael J. Sherratt, Helen K. Graham, Cay M. Kielty, and David F. Holmes. Part III. Molecular Biology of Extracellular Matrix Genes. Screening for Mutations in Cartilage ECM Genes, Michael D. Briggs. Tissue-Specific KO of ECM Proteins, Emilio Hirsch, Mara Brancaccio, and Fiorella Altruda. Homologous Gene Targeting to Study ECM Assembly, Francesco Ramirez, Friedrich Laub, and Hideaki Sumiyoshi. Enhancer Analysis of the a1(II) and a2(XI) Collagen Genes in Transfected Chondrocytes and Transgenic Mice, Noriyuki Tsumaki, Ying Liu, Yoshihiko Yamada, and Paul Krebsbach. Retroviral Delivery of ECM Genes to Cells, Kyonggeun Yoon and Vitali Alexeev. Part IV. Cell Biology of the Extracellular Matrix. Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Subtrates, Christopher S. Chen, Emanuele Ostuni, George M. Whitesides, and Donald E. Ingber. Methods for Preparing Extracellular Matrix and Quantifying Insulin-like Growth Factor-Binding Protein Binding to the ECM, Bo Zheng and David R. Clemmons. Measuring Interactions Between ECM and TGFb-Like Proteins, Sarah L. Dallas. Using Organotypic Tissue Slices as Substrata for the Culture of Dissociated Cells, Daniel E. Emerling and Arthur D. Lander. Neuroepithelial Differentiation Induced by ECM Molecules, José María Frade and Alfred Rodriguez Tébar. Migration Assays for Oligodendrocyte Precursor Cells, Emma E. Frost, Richard Milner, and Charles ffrench-Constant. Cell Adhesion Assays, Martin J. Humphries. Tissue Engineering and Cell-Populated Collagen Matrices, Paul D. Kemp. Solid Phase Assays for Studying ECM Protein-Protein Interactions, A. Paul Mould. Tissue Engineering of Cartilage, Ronda E. Schreiber and Anthony Ratcliffe. Tissue Recombinants to Study Extracellular Matrix Targeting-Basement Membranes, Patricia Simon-Assmann and Michèle Kedinger. Fluorescence Assays to Study Cell Adhesion and Migration In Vitro, Paola Spessotto, Emiliana Giacomello, and Roberto Perris. Analyzing Cell-ECM Interactions in Adult Mammary Gland by Transplantation of Embryonic Mammary Tissue from Knockout Mice, Teresa C. M. Klinowska and Charles H. Streuli. Index.
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