A general method for the detection and mapping of submicrogram quantities of glycosaminoglycan oligosaccharides on polyacrylamide gels by sequential staining with azure A and ammoniacal silver.
AffiliationCancer Research Campaign Department of Medical Oncology, Christie Hospital, Manchester, United Kingdom.
MetadataShow full item record
AbstractA sensitive method has been developed for the visualization of nonradiolabeled glycosaminoglycan oligosaccharides resolved by polyacrylamide gel electrophoresis using fixation with azure A followed by staining with ammoniacal silver. This method, which can detect as little as 1-2 ng of a single oligosaccharide species, can be used to stain a few micrograms of a complex oligosaccharide mixture. The combination of gradient polyacrylamide gel electrophoresis and sequential azure A/silver staining can be applied to the analysis of all the complex glycosaminoglycans (i.e., heparin, heparan sulfate, chondroitin/dermatan sulfate, keratan sulfate) and hyaluronate, as well as to comparisons of specificities of the glycosaminoglycan-degrading enzymes. This procedure may be particularly valuable in situations where the availability of glycosaminoglycan is very limited and/or where radiolabeling is impractical or undesirable.
CitationA general method for the detection and mapping of submicrogram quantities of glycosaminoglycan oligosaccharides on polyacrylamide gels by sequential staining with azure A and ammoniacal silver. 1990, 185 (1):63-70 Anal. Biochem.
- Analysis of glycosaminoglycan-derived oligosaccharides using reversed-phase ion-pairing and ion-exchange chromatography with suppressed conductivity detection.
- Authors: Linhardt RJ, Gu KN, Loganathan D, Carter SR
- Issue date: 1989 Sep
- Quantification and characterization of glycosaminoglycans at the nanogram level by a combined azure A-silver staining in agarose gels.
- Authors: van de Lest CH, Versteeg EM, Veerkamp JH, van Kuppevelt TH
- Issue date: 1994 Sep
- Electrophoresis and detection of nanogram quantities of exogenous and endogenous glycosaminoglycans in biological fluids.
- Authors: al-Hakim A, Linhardt RJ
- Issue date: 1991
- Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycans in sodium dodecyl sulfate-polyacrylamide gels.
- Authors: Møller HJ, Heinegård D, Poulsen JH
- Issue date: 1993 Feb 15
- Cell-associated glycosaminoglycans of human teratocarcinoma-derived cells of line PA 1.
- Authors: Rasilo ML, Renkonen O
- Issue date: 1982 Apr 1