Analysis of fluorophore-labelled hyaluronan and chondroitin sulfate disaccharides in biological samples.
In this report, we describe a polyacrylamide gel electrophoresis for the analysis of fluorophore-labelled hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. The method utilizes derivatization of reducing ends of hyaluronan and the variously sulfated chondroitin sulfate Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis in Tris-HCl buffer (pH 8.8), and polyacrylamide gel (T 25%/C 3.75%). The method was applied to the analysis of GAGs secreted into the culture medium of human aortic smooth muscle cells and the obtained results were compared with those analysed by fluorophore assisted carbohydrate electrophoresis (FACE). The described method is a useful and sensitive tool for the rapid monitoring of GAGs in high number of biologic samples.
Karousou
Evgenia G
EG
Department of Experimental and Clinical Biomedical Sciences, University of Insubria, Varese, Italy.
Porta
Giovanni
G
De Luca
Giancarlo
G
Passi
Alberto
A
D000609:Aminoacridines / Q000737:chemistry; D001011:Aorta / Q000737:chemistry / Q000166:cytology; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000737:chemistry*; D004187:Disaccharides / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006025:Glycosaminoglycans / Q000737:chemistry; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D007558:Italy
D000609:Aminoacridines; D004187:Disaccharides; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; C072515:2-aminoacridone; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates