Abstract
Here, we describe a simple and sensitive method that allows fluorescent detection of glycoproteins on polyvinylidene difluoride (PVDF) membrane. We used periodic acid oxidation of carbohydrate chains of glycoproteins and fluorescent labeling with 8-aminonaphthalene-1,3,6-trisulfonate (AN-TS) by reductive amination. We developed an additional method to enhance the ability of PVDF to absorb glycoproteins by using non-glycoprotein lectin, such as wheat germ agglutinin (WGA), as a link between the PVDF membrane and glycoproteins, resulting in considerably increased detection sensitivity to glycoproteins.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
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Technical Report
MeSH terms
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Absorption
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Animals
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Blotting, Western
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Electrophoresis, Polyacrylamide Gel
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Fluorescent Dyes / analysis*
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Fluorometry / economics
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Fluorometry / methods*
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Glycoproteins / analysis*
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Glycosylation
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Humans
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Immunoblotting
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Lectins / chemistry
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Membranes, Artificial*
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Naphthalenes / analysis*
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Oxidation-Reduction
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Periodic Acid / chemistry
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Polyvinyls / chemistry*
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Protein Processing, Post-Translational
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Sensitivity and Specificity
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Staining and Labeling / economics
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Staining and Labeling / methods*
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Wheat Germ Agglutinins / chemistry
Substances
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Fluorescent Dyes
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Glycoproteins
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Lectins
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Membranes, Artificial
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Naphthalenes
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Polyvinyls
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Wheat Germ Agglutinins
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Periodic Acid
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polyvinylidene fluoride
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8-amino-1,3,6-naphthalenetrisulfonic acid