caa a22 4500 465734227 CHVBK 20180323111750.0 cr unu---uuuuu 170327e19900101xx s 000 0 eng 10.1007/BF01050403 doi (NATIONALLICENCE)springer-10.1007/BF01050403 Description and application of an immunological detection system for analyzing glycoproteins on blots [Elektronische Daten] [Anton Haselbeck, Wolfgang Hösel] 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. Glycoconjugate Journal, 1990 glycoproteins nationallicence carbohydrate structure nationallicence immunochemical methods nationallicence sugar analysis nationallicence gel electrophoresis nationallicence proteins nationallicence Western blotting nationallicence glycosidases nationallicence CpY : Carboxypeptidase Y nationallicence CHO : Chinese hamster ovary nationallicence Dig-hydrazide : digoxigenin-succinyl-ε-caproic acid hydrazide nationallicence 〈Dig〉Fab-AP : polyclonal sheep anti-digoxigenin Fab fragments, conjugated with alkaline phosphatase nationallicence NBT : nitroblue tetrazolium chloride nationallicence SDS-PAGE : sodium dodecylsulfate-polyacrylamide gel electrophoresis nationallicence TCA : trichloroacetic acid nationallicence X-phosphate : 5-bromo-4-chloro-3-indolyl phosphate, 4-toluidine salt nationallicence Haselbeck Anton Biochemica Research Center, Boehringer Mannheim GmbH, D-8132, Tutzing/Obb., Federal Republic of Germany aut Hösel Wolfgang Biochemica Research Center, Boehringer Mannheim GmbH, D-8132, Tutzing/Obb., Federal Republic of Germany aut Glycoconjugate Journal Kluwer Academic Publishers 7/1(1990-01-01), 63-74 0282-0080 7:1<63 1990 7 10719 https://doi.org/10.1007/BF01050403 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01050403 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Haselbeck Anton Biochemica Research Center, Boehringer Mannheim GmbH, D-8132, Tutzing/Obb., Federal Republic of Germany aut NATIONALLICENCE 700 1- Hösel Wolfgang Biochemica Research Center, Boehringer Mannheim GmbH, D-8132, Tutzing/Obb., Federal Republic of Germany aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 7/1(1990-01-01), 63-74 0282-0080 7:1<63 1990 7 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer