| Literature DB >> 2742128 |
V Monboisse1, J C Monboisse, J P Borel, A Randoux.
Abstract
A method for the evaluation of collagen concentrations in the medium of fibroblasts in culture was developed. Collagen was precipitated with other proteins by addition of ethanol and hydrolyzed by 6 M HCl. The primary amino acids of the hydrolyzate were reacted with o-phthalaldehyde (OPA) and secondary amino acids (Pro, Hyp) were derivatized with 9-fluorenylmethyl-chloroformate (FMOC-Cl). The mixture was separated by isocratic HPLC on a reverse-phase column. FMOC-derivatives were detected by fluorometry, whereas OPA-derivatives were not. This method is suitable for the monitoring of collagen metabolism in fibroblast cultures exposed to various effectors.Entities:
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Year: 1989 PMID: 2742128 DOI: 10.1016/0003-2697(89)90330-8
Source DB: PubMed Journal: Anal Biochem ISSN: 0003-2697 Impact factor: 3.365