PMID: 7529008Sep 1, 1994Paper

Quantification and characterization of glycosaminoglycans at the nanogram level by a combined azure A-silver staining in agarose gels

Analytical Biochemistry
C H van de LestT H van Kuppevelt

Abstract

A rapid, sensitive, and nonradioactive method has been developed for the quantification and characterization of glycosaminoglycans. The method is based on the separation of different types of glycosaminoglycans in agarose gel and subsequent fixation and staining with the cationic dye azure A, followed by silver enhancement. Densitometric analysis of the silver deposition gives a linear response between 1 and 20 ng for chondroitin and dermatan sulfate and between 2 and 40 ng for heparan sulfate. The detection limit is about 250 pg. The staining procedure was applied for the quantification and characterization of glycosaminoglycans in human serum, urine, and lung and in mouse kidney glomeruli. It requires only 10 microliters serum, 2 microliters urine, and only a single cryosection in case of tissue. The method is at least as sensitive as staining with radioactive markers and about 200 times more sensitive than conventional glycosaminoglycan-staining methods.

Citations

Oct 15, 2005·Bulletin of Experimental Biology and Medicine·T V RusovaD S Tkachev
Oct 3, 2007·Glycoconjugate Journal·Tessa J WijnhovenToin H van Kuppevelt
Apr 11, 2009·Histochemistry and Cell Biology·Gerdy B Ten DamToin H van Kuppevelt
Oct 9, 2013·BioMed Research International·A M ZaidmanM V Mikhailovsky
Aug 5, 2003·American Journal of Respiratory Cell and Molecular Biology·Nicole C SmitsToin H van Kuppevelt
Dec 24, 2005·The Journal of Biological Chemistry·Gerdy B Ten DamToin H van Kuppevelt
Mar 8, 2011·Tissue Engineering. Part C, Methods·Gerwen LammersWilleke F Daamen
Sep 1, 2015·PloS One·Joost F M LensenAngelique L W M M Rops
Aug 6, 1999·Gene Therapy·F ZhangJ Chroboczek
Nov 16, 2007·Journal of the American Society of Nephrology : JASN·Tessa J M WijnhovenToin H van Kuppevelt
Jan 8, 2016·Scientific Reports·Corien OostendorpToin H van Kuppevelt
Nov 30, 2016·PloS One·Jasper J van GemstJohan van der Vlag
Jan 26, 2007·Journal of the American Society of Nephrology : JASN·Tessa J M WijnhovenToin H van Kuppevelt
Sep 23, 1997·Analytical Biochemistry·T ToidaR J Linhardt
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·H Savolainen
Sep 8, 2018·Molecular Therapy. Methods & Clinical Development·Haiyan FuJoseph Muenzer
Mar 15, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Nicola Volpi, Francesca Maccari
May 4, 2010·Experimental Cell Research·Xander M R van WijkToin H van Kuppevelt

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