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A rapid method for fractionation and purification of a 92 kDa cartilage glycoprotein
Authors:Roussidis Andreas E  Anagnostides Stavros T
Institution:Department of Chemistry, Section of Organic Chemistry, Biochemistry and Natural Products, Laboratory of Biochemistry, University of Patras, 261 10 Patras, Greece.
Abstract:Matrix glycoproteins are among the main components that contribute to the properties of cartilage. In this article we report on the development of a rapid method for the fractionation and purification of a 92 kDa glycoprotein from chick sternal cartilage. The developed procedure involves ion-exchange chromatography on DEAE-Sephacel, gel permeation chromatography on Sepharose CL-6B and semi-preparative SDS-polyacrylamide gel electrophoresis. Identification of protein was performed by western blotting using specific antibodies and purity by capillary electrophoresis. The proposed method is superior to those previously published since it eliminates the step of density gradient centrifugation.
Keywords:matrix glycoproteins  ion‐exchange chromatography  gel permeation chromatography  polyacrylamide gel electrophoresis  capillary electrophoresis
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