title: Solid-phase assay of lectin activity using HRP-conjugated glycoproteins.
abstract: Various enzyme-conjugated probes have been widely used for detection of specific interactions between biomolecules. In the case of glycan-protein interaction, horseradish peroxidase (HRP)-conjugated glycoproteins (HRP-GPs) are useful for the detection of carbohydrate-binding activity of plant and animal lectins. In this chapter, a typical solid-phase assay of the carbohydrate-binding activity of Sophora japonica agglutinin I, a Gal/GalNAc-specific lectin, using HRP-conjugated asialofetuin is described. HRP-GPs are versatile tools for probing lectin activities in crude extracts, screening many samples at one time, and applicable not only for solid-phase binding assays but also samples which are dot- or Western-blotted onto the membrane.
authors.affiliation: Division of Natural and Applied Sciences, Graduate School of Humanities and Sciences, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo, 112-8610, Japan, aikawa.kyoko@ocha.ac.jp.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Kojima-Aikawa
mesh_terms: D001681:Biological Assay / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism*; D006385:Hemagglutination Inhibition Tests; D006735:Horseradish Peroxidase / Q000378:metabolism*; D015335:Molecular Probes / Q000378:metabolism; D024301:Plant Bark / Q000737:chemistry; D037121:Plant Lectins / Q000378:metabolism*; D029909:Sophora / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D015335:Molecular Probes; D037121:Plant Lectins; D006735:Horseradish Peroxidase