DESCRIPTION AND APPLICATION OF AN IMMUNOLOGICAL DETECTION SYSTEM FOR ANALYZING GLYCOPROTEINS ON BLOTS

被引:97
作者
HASELBECK, A
HOSEL, W
机构
[1] Biochemica Research Center, Boehringer Mannheim GmbH, Tutzing/Obb.
关键词
carbohydrate structure; gel electrophoresis; glycoproteins; glycosidases; immunochemical methods; proteins; sugar analysis; Western blotting;
D O I
10.1007/BF01050403
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
By introducing the steroid hapten digoxigenin specifically into sugars, a sensitive detection system for glycoproteins on blots has been developed. Sugars are oxidized to obtain aldehyde groups, which then react with digoxigenin-succinyl-ε-amido caproic acid hydrazide. A high-affinity antibody, conjugated to alkaline phosphatase, is used for the detection of the incorporated digoxigenin. This system allows the detection of nanogram-amounts of glycoproteins on blots, and it's specificity allows a clear distinction of a glycoprotein from a non-glycoprotein. In combination with endo- and exoglycosidases it is very useful for determining the type of carbohydrate linkage in a glycoprotein, and by varying the oxidation conditions, specific labeling of sialic acids and terminal galactoses can be achieved. © 1990 Glycoconjugate Journal.
引用
收藏
页码:63 / 74
页数:12
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