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The study of the extracellular matrix has significantly expanded over the past decade. Although thoroughly researched, the complexity and the dynamic nature of the matrix are still under extensive investigation, yielding a growing number of methods of analysis.
In Extracellular Matrix Protocols: Second Edition, expert researchers explore this developing field and provide a number of techniques which can be reproduced in most cell biology labs. This edition contains methods that represent cutting edge unique technologies which are well worth exposure to the broad field and may encourage collaboration with expert labs. Chapters include information on molecular biology techniques to study gene expression, biophysical and biochemical methods for the analysis of structure and composition, and cell biology methods for assessment of cell behavior, matrix assembly and tissue engineering applications.
Composed in the highly successful Methods in Molecular Biology(tm) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Extracellular Matrix Protocols: Second Edition investigates the diverse roles of the extracellular matrix, providing important tools for all researchers in the ECM community.
Preface
Contributors
I. Molecular Biology
Retroviral delivery of ECM genes
Vitali Alexeev and Olga Igoucheva
Tissue-Specific KO of ECM Proteins
Mara Brancaccio, Emila Turco and Emilio Hirsch
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
Tissue Recombinants to Study Extracellular Matrix Targeting to Basement Membranes
Patricia Simon-Assmann, Anne-Laure Bolcato-Bellemin, Annick Klein and Mich#le Kedinger
Preparation of recombinant fibronectin fragments for functional and structural studies
David Staunton, Christopher J Millard, Radu Aricescu and Iain D Campbell
II. Biochemical and biophysical analysis
Quantitative Determination of Collagen Cross-links
Nicholas C Avery, Trevor J Sims and Allen J Bailey
ECM Macromolecules: Height-mapping and Nano-mechanics Using Atomic Force Microscopy
Nigel W. Hodson, Cay M. Kielty and Michael J. Sherratt
Atomic Force Microscopy Measurements of Intermolecular Binding Forces-
Gradimir N. Misevic, Yannis Karamanos and Nikola J. Misevic
Mass-Mapping of ECM Macromolecules by Scanning Transmission Electron Microscopy
Michael J. Sherratt, Helen K. Graham, Cay M. Kielty and David F. Holmes
Chemical Microscopy of Biological Samples by Dynamic Mode Secondary Ion Mass Spectrometry (SIMS)
Gradimir N. Misevic, Bernard Rasser, Vic Norris, C,rLeiaer olGaL and Camille Ripoll
ECM Macromolecules: Rotary Shadowing and Transmission Electron Microscopy
Michael J. Sherratt, Roger S. Meadows, Helen K. Graham, Cay M. Kielty and David F. Holmes
Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Substrates
Emanuele Ostuni, George M. Whitesides, Donald E. Ingber and Christopher S. Chen
Solid Phase Assays for Studying ECM Protein-Protein Interactions
Paul A Mould
III. Cell biology assays
Cell Adhesion Assays
Martin J. Humphries
ECM Degradation Assays for Analyzing Local Cell Invasion
Vira V. Artym, Kenneth M. Yamada, Susette C. Mueller
Fluorescence-Based Assays for In Vitro Analysis of Cell Adhesion and Migration
Paola Spessotto, Katia Lacrima, Pier Andrea Nicolosi, Eliana Pivetta, Martina Scapolan and Roberto Perris
Fibrin Gel Model for Assessment of Cellular Contractility
Sharona Even-Ram
Fluorescent labeling techniques for investigation of fibronectin fibrillogenesis ( Labeling fibronectin fibrillogenesis)
Roumen Pankov and Albena Momchilova
Stromagenesis During Tumorigenesis: Characterization of Tumor-associated Fibroblasts and Stroma-derived 3D Matrices
Remedios Castell-Cros and Edna Cukierman
IV. Organ models
ECM and FGF-dependent assay of embryonic SMG epithelial morphogenesis: investigating growth factor /matrix regulation of gene expression during submandibular gland development
Ivan T. Rebustini and Matthew P. Hoffman
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
V. Tissue Engineering
Characterizing ECM Production by Cells Encapsulated in Hydrogels
Iossif A. Strehin and Jennifer H. Elisseeff
Tissue Engineering and Cell-Populated Collagen Matrices Paul D. Kemp
