Centrifugal ultrafiltration-HPLC method for interaction analysis between lectins and sugars
The centrifugal ultrafiltration-HPLC method is a simple and rapid method for analyzing the binding interaction between lectins and sugars (oligosaccharides). In this method, a lectin is mixed with a fluorescent-labeled oligosaccharide in buffer and the unbound oligosaccharide recovered by centrifuga...
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Published in | Methods in molecular biology (Clifton, N.J.) Vol. 1200; p. 173 |
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Main Authors | , |
Format | Journal Article |
Language | English |
Published |
United States
2014
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Abstract | The centrifugal ultrafiltration-HPLC method is a simple and rapid method for analyzing the binding interaction between lectins and sugars (oligosaccharides). In this method, a lectin is mixed with a fluorescent-labeled oligosaccharide in buffer and the unbound oligosaccharide recovered by centrifugal ultrafiltration is isolated and quantified by high-performance liquid chromatography. The binding activity is defined as a ratio (percentage) of the amount of bound oligosaccharide to that added, where the former is obtained by subtracting the amount of unbound oligosaccharide from the latter. The oligosaccharide-binding specificity of a lectin can be determined by comparing the binding activities with a variety of fluorescent-labeled oligosaccharides. The association constant and the optimum pH and temperature of the binding interaction between lectins and fluorescent-labeled oligosaccharides can be easily analyzed by this method. |
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AbstractList | The centrifugal ultrafiltration-HPLC method is a simple and rapid method for analyzing the binding interaction between lectins and sugars (oligosaccharides). In this method, a lectin is mixed with a fluorescent-labeled oligosaccharide in buffer and the unbound oligosaccharide recovered by centrifugal ultrafiltration is isolated and quantified by high-performance liquid chromatography. The binding activity is defined as a ratio (percentage) of the amount of bound oligosaccharide to that added, where the former is obtained by subtracting the amount of unbound oligosaccharide from the latter. The oligosaccharide-binding specificity of a lectin can be determined by comparing the binding activities with a variety of fluorescent-labeled oligosaccharides. The association constant and the optimum pH and temperature of the binding interaction between lectins and fluorescent-labeled oligosaccharides can be easily analyzed by this method. |
Author | Hirayama, Makoto Hori, Kanji |
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SubjectTerms | Amination Centrifugation - methods Chromatography, High Pressure Liquid - methods Fluorescent Dyes - chemistry Fluorometry Kinetics Lectins - metabolism Oligosaccharides - chemistry Oligosaccharides - isolation & purification Oligosaccharides - metabolism Protein Binding Substrate Specificity Temperature Ultrafiltration - methods |
Title | Centrifugal ultrafiltration-HPLC method for interaction analysis between lectins and sugars |
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