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Increased selectivity in the detection of glycoproteins on nitrocellulose membranes by washing with sodium hydroxide solution
Author(s) -
Kondo Maki,
Harada Hiroshi,
Sunada Saburo,
Yamaguchi Takashi
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150120918
Subject(s) - nitrocellulose , chemistry , glycoprotein , membrane , sodium hydroxide , chromatography , biotin , sodium periodate , biochemistry , bovine serum albumin , selectivity , horseradish peroxidase , organic chemistry , enzyme , catalysis
Abstract We increased selectivity in the detection of glycoproteins on nitrocellulose membranes by introducing a washing step using sodium hydroxide solution. Glycoproteins on the nitrocellulose membrane were first oxidized by sodium periodate; biotin hydrazide was then coupled to the aldehyde groups generated in the sugar moiety of the glycoproteins. The membrane was washed twice using sodium hydroxide solution, and avidin‐horseradish peroxidase was then coupled to the remaining biotin. This system allows the detection of nanograms of glycoproteins on nitrocellulose membranes, and its specificity allows the clear distinction of glycoproteins from the nonglycosylated protein of bovine serum albumin.