FLUOROMETRIC ASSAY OF O-LINKED GLYCOPROTEINS BY REACTION WITH 2-CYANOACETAMIDE
FLUOROMETRIC ASSAY OF O-LINKED GLYCOPROTEINS BY REACTION WITH 2-CYANOACETAMIDE
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DOI：
10.1016/0003-2697(87)90108-4
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发表时间：
1987-05-15
影响因子：
2.9
通讯作者：
WETMORE, RF
中科院分区：
文献类型：
--
作者：
CROWTHER, RS;WETMORE, RF
o -糖基化糖蛋白的一种简单测定方法，涉及低聚糖的. β释放。报道了用稀碱消除和随后用2-氰乙酰胺衍生化还原端的反应。该方法可在30min内定量测定粘蛋白的微克量。该试验对蛋白质或n -连接糖蛋白的敏感性比对犬气管黏液相应重量的敏感性低30倍，对DNA的敏感性低100倍。在测试的物质中，只有氯化铯和硫氰酸钾会造成严重干扰，但这两种情况都不足以严重到阻止该方法用于监测利用这些试剂的粘蛋白纯化方案。犬气管粘蛋白重复分析的变异系数分别为14.0、5.0和2.0 .mu。G)分别为3.6、6.5和12.3%。
A simple assay for O-glycosylated glycoproteins involving the liberation of oligosaccharides by .beta.-elimination with dilute alkali and the subsequent derivatization of the reducing end with 2-cyanoacetamide is reported. The method can be used to quantitate microgram amounts of mucin within 30 min. The assay is 30 times less sensitive to protein or N-linked glycoproteins and 100 times less sensitive to DNA than to the corresponding weight of canine tracheal mucin. Of the substances tested, only cesium chloride and potassium thiocyanate caused substantial interference, but in neither case was this sufficiently serious to prevent the method being used for monitoring mucin purification schemes utilizing these reagents. The coefficients of variation for replicate analyses of canine tracheal mucin (14.0, 5.0, and 2.0 .mu.g) were 3.6, 6.5, and 12.3%, respectively.