Adherence of Glycosaminoglycans to Calcium-Oxalate Crystals
Calcium oxalate (CaOx) crystals were manufactured in a 10-mM Ca++ excess solution by combining 5.0 ml of 0.3-M K-oxalate, 1.0 ml of 1.65-M CaCl2, and 1.0 ml of 1.5-M Na-cacodylate buffer (pH 7.3, 20°C) in borosilicate glass vials; the mixture was agitated for 30 min in a thermostatically-controlled shaker bath (37°C). The macromolecules were previously dissolved in 0.3-M KC1. The mixture was placed back in the water bath for 2 h, centrifuged at 200 g for five min, and the supernatant was filtered through 0.2-μm polycarbonate filters. The equilibrium concentration of the adsorbate was determined by measuring the concentration of glycosaminoglycan (GAG) (uronic acid) using the solution-depletion method.
KeywordsHyaluronic Acid Heparan Sulfate Chondroitin Sulfate Calcium Oxalate Dermatan Sulfate
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