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Methods Cell Biol. 2018;143:133-146. doi: 10.1016/bs.mcb.2017.08.008. Epub 2017 Nov 6.

Measurement of elastin, collagen, and total protein levels in tissues.

Author information

1
Washington University School of Medicine, St. Louis, MO, United States.
2
The University of Texas Health Science Center, Tyler, TX, United States.
3
Washington University School of Medicine, St. Louis, MO, United States. Electronic address: tbroekel@wustl.edu.

Abstract

Elastin and collagen levels in tissues are frequently difficult to measure because of each protein's limited solubility. This chapter provides detailed methodology for the determination of elastin, collagen, and total protein levels in a single tissue sample. All three assays start with an acid hydrolysate of the tissue, which breaks the tissue-associated proteins down to their component amino acids. Marker amino acids unique to each protein (desmosine for elastin and hydroxyproline for collagen) are then quantified. Total protein content, useful as a denominator for data normalization, can also be measured from a portion of the hydrolysate using an assay for free amino groups. These measurements are performed using convenient 96-well assay plates and require only a plate reader to determine absorbance.

KEYWORDS:

Collagen; Desmosine; ELISA; Elastin; Hydroxyproline

PMID:
29310774
DOI:
10.1016/bs.mcb.2017.08.008
[Indexed for MEDLINE]

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