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Quantification of collagen protein

AZ Ariella Zehender
JH Jingang Huang
AG Andrea-Hermina Györfi
AM Alexandru-Emil Matei
TT Thuong Trinh-Minh
XX Xiaohan Xu
YL Yi-Nan Li
CC Chih-Wei Chen
JL Jianping Lin
CD Clara Dees
CB Christian Beyer
KG Kolja Gelse
ZZ Zhong-Yin Zhang
CB Christina Bergmann
AR Andreas Ramming
WB Walter Birchmeier
OD Oliver Distler
GS Georg Schett
JD Jörg H. W. Distler
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The amount of soluble collagen in cell culture supernatants was quantified using the SirCol collagen assay (Biocolor, Belfast, Northern Ireland). The total collagen content of tissue samples was determined by hydroxyproline assays using the chloramines-T method28,63. In brief, samples were digested with 6 M HCl for 4–6 h. Samples were centrifuged to remove debris and pH of the solution is adjusted to 7. Samples were hydrolyzed by incubation at 60 °C for 30 min. The cloramines-T was added to the hydrolyzate to allow oxidation followed by the addition of Ehrlich’s aldehyde reagent. The absorbance intensity of each sample was analyzed at 550 nm using a microtiter plate reader spectrophotometer.

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