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Matrix metalloproteinase protocols / edited by Ian M. Clark ; consulting editors, David A. Young, Andrew D. Rowan. — 2nd ed. — New York, N.Y. : Humana Press, c2010. – (58.17/M592/v.622) |
Contents
Contents
Contributors
SECTION I MMPS AND TIMPS: AN OVERVIEW 1
1. Metalloproteases and the Degradome
2. Mouse Models of MMP and TIMP Function
SECTION II EXPRESSION AND PURIFICATION OF MMPS AND TIMPs 53
3. Expression of Recombinant MMP-28 in Mammalian Cells 55
4. Expression of Recombinant Matrix Metalloproteinases in Escherichia coli
5. Expression of Recombinant ADAMTS in Insect Cells 83
6. Expression and Purification of Membrane-Type MMPs
7. Refolding of TIMP-2 from Escherichia coli Inclusion Bodies
8. Purification of MMPs and TIMPs
SECTION III DETECTION OF MMPS AND TIMPS 157
9. Real-Time PCR Expression Profiling of MMPs and TIMPs
10. Analysis of the Degradome with the CLIP-CHIPTM Microarray 175
11. In Situ Hybridization for Metalloproteinases and Their Inhibitors 195
12. Immunohistochemistry of MMPs and TIMPs
13. Single Nucleotide Polymorphism Genotyping in MMP Genes: The 5' Nuclease Assay
SECTION IV ASSAY OF MMP AND TIMP ACTIVITIES 231
14. Methods for Studying Activation of Matrix Metalloproteinases 233
15. Assay of Matrix Metalloproteinases Against Matrix Substrates
16. Zymography and Reverse Zymography for Detecting MMPs and TIMPs 257
17. In Situ Zymography 271
18. Near-Infrared Optical Proteolytic Beacons for In Vivo Imaging of Matrix Metalloproteinase Activity 279
19. Neoepitope Antibodies Against MMP-Cleaved and Aggrecanase-Cleaved Aggrecan
20. In Vitro Model of Cartilage Degradation
21. Immunoassays for Collagenase-Mediated Cleavage of Type I and II Collagens
22. Collagen Degradation Assays
23. Analysis of MMP-Dependent Cell Migration and Invasion 379
24. Using Fluorogenic Peptide Substrates to Assay Matrix Metalloproteinases 393
25. Kinetic Analysis of the Inhibition of Matrix Metalloproteinases: Lessons from the Study of Tissue Inhibitors of Metalloproteinases 435
26. Identification of Cellular MMP Substrates Using Quantitative Proteomics: Isotope-Coded Affinity Tags (ICAT) and Isobaric Tags for Relative and Absolute Quantification (iTRAQ) 451
27. Mechanism-Based Profiling of MMPs 471
Subject Index 489
