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9781588299840

Extracellular Matrix Protocols

by ;
  • ISBN13:

    9781588299840

  • ISBN10:

    1588299848

  • Edition: 2nd
  • Format: Hardcover
  • Copyright: 2009-07-04
  • Publisher: Humana Pr Inc

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Summary

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 BiologyTM 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.

Table of Contents

Prefacep. v
Contributorsp. ix
Molecular Biology
Retroviral Delivery of ECM Genesp. 3
Tissue-Specific KO of ECM Proteinsp. 15
Recombinant Collagen Trimers from Insect Cells and Yeastp. 51
Eukaryotic Expression and Purification of Recombinant Extracellular Matrix Proteins Carrying the Strep II Tagp. 63
Preparation of Recombinant Fibronectin Fragments for Functional and Structural Studiesp. 73
Biochemical and Biophysical Analysis
Quantitative Determination of Collagen Cross-linksp. 103
ECM Macromolecules: Height-Mapping and Nano-Mechanics Using Atomic Force Microscopyp. 123
Atomic Force Microscopy Measurements of Intermolecular Binding Forcesp. 143
Mass-Mapping of ECM Macromolecules by Scanning Transmission Electron Microscopyp. 151
Chemical Microscopy of Biological Samples by Dynamic Mode Secondary Ion Mass Spectrometryp. 163
ECM Macromolecules: Rotary Shadowing and Transmission Electron Microscopyp. 175
Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Substratesp. 183
Solid Phase Assays for Studying ECM Protein-Protein Interactionsp. 195
Cell Biology Assays
Cell Adhesion Assaysp. 203
ECM Degradation Assays for Analyzing Local Cell Invasionp. 211
Fluorescence-Based Assays for In Vitro Analysis of Cell Adhesion and Migrationp. 221
Fibrin Gel Model for Assessment of Cellular Contractilityp. 251
Fluorescent Labeling Techniques for Investigation of Fibronectin Fibrillogenesis (Labeling Fibronectin Fibrillogenesis)p. 261
Stromagenesis During Tumorigenesis: Characterization of Tumor-Associated Fibroblasts and Stroma-Derived 3D Matricesp. 275
Organ Models
Tissue Recombinants to Study Extracellular Matrix Targeting to Basement Membranesp. 309
ECM and FGF-Dependent Assay of Embryonic SMG Epithelial Morphogenesis: Investigating Growth Factor/Matrix Regulation of Gene Expression During Submandibular Gland Developmentp. 319
Analyzing how Cell Adhesion Controls Mammary Gland Function by Transplantation of Embryonic Mammary Tissue from Knockout Micep. 331
Tissue Engineering
Characterizing ECM Production by Cells Encapsulated in Hydrogelsp. 349
Tissue Engineering and Cell-Populated Collagen Matricesp. 363
Indexp. 371
Table of Contents provided by Ingram. All Rights Reserved.

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