Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.actbio.2010.02.004
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dc.titleEssential modification of the Sircol Collagen assay for the accurate quantification of collagen content in complex protein solutions
dc.contributor.authorLareu, R.R.
dc.contributor.authorZeugolis, D.I.
dc.contributor.authorAbu-Rub, M.
dc.contributor.authorPandit, A.
dc.contributor.authorRaghunath, M.
dc.date.accessioned2014-06-17T09:43:43Z
dc.date.available2014-06-17T09:43:43Z
dc.date.issued2010-08
dc.identifier.citationLareu, R.R., Zeugolis, D.I., Abu-Rub, M., Pandit, A., Raghunath, M. (2010-08). Essential modification of the Sircol Collagen assay for the accurate quantification of collagen content in complex protein solutions. Acta Biomaterialia 6 (8) : 3146-3151. ScholarBank@NUS Repository. https://doi.org/10.1016/j.actbio.2010.02.004
dc.identifier.issn17427061
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/67041
dc.description.abstractCollagen contains the unique imino acid hydroxyproline (HyPro), which is involved in the stabilization of this triple helical molecule. The concentration of HyPro is customarily used to calculate the total collagen content in a cell culture environment and in acid hydrolysates of normal and pathophysiological tissues. Radiolabelling, chromatographic and calorimetric assays have been developed over the years for the accurate determination of collagen content through HyPro estimation. Recently, the Sircol Collagen Assay (SCA) has been almost exclusively adopted as the fastest and simplest colorimetric method for the determination of collagen concentration in complex protein solutions. We show here that the colorimetric SCA, which is based on the binding of Sirius red (SR) to collagen, is flawed by interference of non-collagenous proteins (e.g. serum). In fact, we demonstrate that SCA in cell culture systems and tissue hydrolysates results in a dramatic overestimation of collagen content ranging from 3- to 24-fold. In order to rescue this otherwise very practical assay, we introduce a simple purification procedure that allows the removal of interfering non-collagenous proteins from culture media and tissue samples so that accurate measurements with SCA are now possible. © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1016/j.actbio.2010.02.004
dc.sourceScopus
dc.subjectCollagen content
dc.subjectHydroxyproline assay
dc.subjectSircol assay
dc.subjectSirius red
dc.subjectUltrafiltration
dc.typeArticle
dc.contributor.departmentBIOENGINEERING
dc.description.doi10.1016/j.actbio.2010.02.004
dc.description.sourcetitleActa Biomaterialia
dc.description.volume6
dc.description.issue8
dc.description.page3146-3151
dc.identifier.isiut000279583400032
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