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Methods for Measuring Type I Collagen Synthesis In Vitro

  • Protocol
Fibrosis Research

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 117))

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Abstract

The excess accumulation of type I collagen within tissues leads to organ dysfunction and occurs as a result of an imbalance between synthesis and degradation. This chapter outlines several methods to assess the in vitro production of type I collagen that are employed in our laboratory. We describe Western immunoblotting of intact α1(I) collagen using antibodies directed to α1(I) collagen amino and carboxyl propeptides. The measurement of α1(I) collagen mRNA levels using real-time polymerase chain reaction is then outlined. Finally, methods to determine the transcriptional regulation of α1(I) collagen using a nuclear run-on assay are described.

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Acknowledgments

This work was supported by the National Institutes of Health grants K08-HL04232 and R01-HL66547 and the VA Research Enhancement Award Program.

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Authors and Affiliations

  1. The Pulmonary Center, Boston University School of Medicine, The Boston VA Medical Center, Boston, MA

    David C. Rishikof, Ping-Ping Kuang, Mangalalaxmy Subramanian & Ronald H. Goldstein

Authors
  1. David C. Rishikof
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  2. Ping-Ping Kuang
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  3. Mangalalaxmy Subramanian
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  4. Ronald H. Goldstein
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Editor information

Editors and Affiliations

  1. Feinberg School of Medicine, Northwestern University, Chicago, IL

    John Varga MD

  2. College of Physicians and Surgeons, Columbia University, New York, NY

    David A. Brenner MD

  3. University of Michigan Medical School, Ann Arbor, MI

    Sem H. Phan MD, PhD

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© 2005 Humana Press Inc.

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Rishikof, D.C., Kuang, PP., Subramanian, M., Goldstein, R.H. (2005). Methods for Measuring Type I Collagen Synthesis In Vitro. In: Varga, J., Brenner, D.A., Phan, S.H. (eds) Fibrosis Research. Methods in Molecular Medicine, vol 117. Humana Press. https://doi.org/10.1385/1-59259-940-0:129

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  • DOI: https://doi.org/10.1385/1-59259-940-0:129

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-479-1

  • Online ISBN: 978-1-59259-940-0

  • eBook Packages: Springer Protocols

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