Isolation and characterization of proteoglycans from different tissues.
Two chromatographic procedures for the isolation and purification of proteoglycans (PG) and their related glycosaminoglycan (GAG) peptides are described. PG from human aorta were isolated from tissue extract by sequential ion-exchange, size-exclusion and hydroxyapatite chromatography. Final purification of samples was achieved by chromatography on Mono Q. Homogeneity of samples was demonstrated by Western blot analysis of biotin-labelled compounds prior to and after enzymatic digestion and dual-wavelength detection in size-exclusion chromatography. The purity of samples obtained by the procedure described was sufficient for protein sequence analysis. GAG preparations of bovine trachea cartilage were purified by the sequential use of strong anion-exchange supports. Molecular weight distribution and sensitivity to treatment with glycan-specific enzymes was shown by size-exclusion chromatography.
Stöcker
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Department of Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Aachen, F.R.G.
Fischer
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D000818:Animals; D001011:Aorta / Q000737:chemistry; D015153:Blotting, Western; D002356:Cartilage / Q000737:chemistry; D002417:Cattle; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D017886:Durapatite; D006025:Glycosaminoglycans / Q000302:isolation & purification; D019763:Heparin Lyase; D006801:Humans; D006882:Hydroxyapatites; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011509:Proteoglycans / Q000302:isolation & purification*; D014132:Trachea / Q000737:chemistry
D006025:Glycosaminoglycans; D006882:Hydroxyapatites; D011509:Proteoglycans; D017886:Durapatite; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase; C021349:heparitinsulfate lyase