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M1563 Sigma

Matrix Metalloproteinase Protocols

  •  ISBN-10 0-89603-733-9

  •  ISBN-13 978-0-89603-733-5

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General description

Experts detail proven laboratory techniques for the study of MMPs. Methods include those for the expression and purification of MMPs and TIMPs, for the detection of MMPs and TIMPs at both the protein and mRNA levels, and for the assay of MMP and TIMP activities in a variety of circumstances. Each method includes step-by-step instructions and notes variant applications and pitfalls to avoid. A selective overview of the MMP arena states where the field has been, where it is, and where it is going.

Table of Contents

Part I. MMPs and TIMPs
An Overview of the Field MMPs and TIMPs: An Historical Perspective, J. Frederick Woessner, Jr.
Strategies for Cloning New MMPs and TIMPs, Gloria Velasco and Carlos López-Otín
Structural Studies of MMPs and TIMPs, Wolfram Bode and Klaus Maskos
Matrix Metalloproteinase Substrate Binding Domains, Modules and Exosites: Overview and Experimental Strategies, Christopher M. Overall
The Matrix Metalloproteinase (MMP) and Tissue Inhibitor of Metalloproteinase (TIMP) Genes: Transcriptional and Posttranscriptional Regulation, Signal Transduction and Cell-Type-Specific Expression, Matthew P. Vincenti.
Models for Gain-of-Function and Loss-of-Function of MMPs: Transgenic and Gene Targeted Mice, Lisa M. Coussens, et.al.
Part II. Expression and Purification of MMPs and TIMPs
Expression of MMPs and TIMPs in Mammalian Cells, Karen M.-L. Yeow, et.al.
Expression of Recombinant Matrix Metalloproteinases in Escherichia coli, L. Jack Windsor and Darin L. Steele
Expression of Human Collagenase I (MMP-1) and TIMP-1 in a Baculovirus-Based Expression System, Rüdiger Vallon and Peter Angel
Expression of Recombinant Matrix Metalloproteinases in Yeast, Glenn A. Doyle
Expression of Recombinant Membrane-Type MMPs, Michael J. Butler, et.al.
Refolding of TIMP-2 from Escherichia coli Inclusion Bodies, Richard A. Williamson
Expression and Refolding of Full-Length Human TIMP-1, Deborah Davis.
Purification of MMPs and TIMPs, Ken-ichi Shimokawa and Hideaki Nagase
Part III. Detection of MMPs and TIMPs
Monitoring MMP and TIMP mRNA Expression by RT-PCR, Howard Wong, et.al.
Measuring Transcription of Metalloproteinase Genes: Nuclear Run-Off Assay vs Analysis of hnRNA, Anne M. Delany
In Situ Hybridization for Metalloproteinases and Their Inhibitors, Tina L. Hurskainen and Suneel S. Apte
Use of EIA to Measure MMPs and TIMPs, Noboru Fujimoto and Kazushi Iwata
Immunohistochemistry of MMPs and TIMPs, Yasunori Okada
Detecting Polymorphisms in MMP Genes, Shu Ye and Adriano M. Henney
Part IV. Assay of MMP and TIMP Activities
Methods for Studying Activation of Matrix Metalloproteinases, Vera Knäuper and Gillian Murphy
Assay of Matrix Metalloproteinases Against Matrix Substrates, Timothy E. Cawston, et.al.
Zymography and Reverse Zymography for Detecting MMPs and TIMPs, Susan P. Hawkes, et.al.
Detection of Focal Proteolysis Using Texas-Red-Gelatin, Rosalind M. Hembry
Antibodies to MMP-Cleaved Aggrecan, Amanda J. Fosang, et.al.
Cartilage Proteoglycan Release Assay, Caron J. Billington
Immunoassay for Collagenase-Mediated Cleavage of Types I and II Collagens, R. Clark Billinghurst, et.al.
Collagen Degradation Assays, Anthony P. Hollander.
Invasion Assays and Matrix Metalloproteinases: Quantification of Cellular Invasion Using Propidium Iodide Labeling and Confocal Laser Scanning Microscopy, Ulrike Benbow, et.al.
Using Fluorogenic Peptide Substrates to Assay Matrix Metalloproteinases, Gregg B. Fields
Kinetic Analysis of the Inhibition of Matrix Metalloproteinases by Tissue Inhibitor of Metalloproteinases (TIMP), Mike Hutton and Frances Willenbrock
Assaying Growth Factor Activity of Tissue Inhibitors of Metalloproteinases, Kyoko Yamashita and Taro Hayakawa.
Index.

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