title: Detection of glycolipids as biotinylated derivatives using enhanced chemiluminescence.
abstract: A new method for the detection of glycolipids as biotinylated derivatives is presented. This method is based on the detection of lipids by enhanced chemiluminescence (ECL) following conjugation with streptavidin-horseradish peroxidase (SHRP). Partial biotinylation of glycolipids is achieved after mild oxidation of the glycan moiety with Na-meta-periodate to increase the availability of biotin-reactive sites. There are several significant advantages of the glycolipid ECL detection: it avoids the need for radio-labeling; it provides the possibility of antibody probing; it allows reprobing of SHRP conjugate to adjust background and luminol light emission levels; it permits easy and accurate quantification of glycolipids at low concentrations; and it involves nondestructive staining, thereby enabling further molecular analysis.
authors.affiliation: John Innes Centre, Norwich, United Kingdom.
authors.forename: L E
authors.initials: LE
authors.lastname: Hernández
authors.forename: N J
authors.initials: NJ
authors.lastname: Brewin
authors.forename: B K
authors.initials: BK
authors.lastname: Drøbak
mesh_terms: D001710:Biotin; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006017:Glycolipids / Q000032:analysis*; D008163:Luminescent Measurements; D009195:Peroxidase
chemical_list: D006017:Glycolipids; D001710:Biotin; D009195:Peroxidase