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Quantitative evaluation of collagen fractions separated by acrylamide gel electrophoresis: its application for
Abstract:
A method for the quantitation of collagen chains or cyanogen bromide peptides separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis is described. After electrophoresis, the bands, slightly stained by Coomassie brilliant blue, are cut and hydrolysed in 6 M hydrochloric acid. Proline and hydroxyproline are measured by a fluorometric procedure after thin-layer separation. When the method is applied to the fractions solubilized by pepsin digestion, it provides a measurement of type I, III, IV and V collagens. When it is applied to cyanogen bromide peptides, its permits the calculation of the proportions of type I and III collagens. Applied to polycystic kidney, this method indicates a significant increase of type I and a limited increase of type IV collagen in this abnormal tissue compared with normal kidney.