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Extracellular Matrix Protocols

Second Edition

Series: Methods in Molecular Biology

by Sharona Even-Ram and Vira Artym

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Main description:

The study of the extracellular matrix (ECM) and its diverse roles in tissue scaffolding and cellular signaling in both physiological and pathological processes has significantly expanded over the past decade. Although well appreciated, the structural and biochemical complexity and the dynamic nature of the living matrix are still under extensive inv- tigation, yielding a growing number of methods with varying degree of sophistication and intricacy. In this edition of Extracellular Matrix Protocols, we compiled a variety of methods that can be readily reproduced in most cell biology laboratories, as well as several cutting-edge technologies that are indeed available in a limited number of centers, but are well worth the awareness and exposure to the ECM research community. As in the previous edition, the book chapters are divided into sections that rep- sent molecular biology techniques to study gene expression, biophysical and biochemical methods for the analysis of structure and composition, cell biology methods for the asse- ment of cell behavior and matrix assembly and tissue engineering applications. All chapters were contributed by scientists who developed the methods or mastered and perfected methods that were routinely used in their laboratories. An effort was made to provide practical working details and helpful notes for the nonexpert user in order to assist reproducibility and accuracy. We hope that these valuable protocols will become helpful tools for ECM researchers and will be further developed and tailored to the s- cific needs of a growing number of applications.

Contents:

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 Michele 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, Cedric Derue, David Gibouin, Fabrice Lefebvre, Marie-Claire Verdus, Anthony Delaune, Guillaume Legent and CamilleRipoll

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 Castello-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

PRODUCT DETAILS

ISBN-13: 9781588299840
Publisher: Springer (Humana Press Inc.)
Publication date: March, 2009
Pages: 334
Weight: 1026g
Availability: Available
Subcategories: Anatomy

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