Detection of glycolipids as biotinylated derivatives using enhanced chemiluminescence.
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.
Hernández
L E
LE
John Innes Centre, Norwich, United Kingdom.
Brewin
N J
NJ
Drøbak
B K
BK
D001710:Biotin; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006017:Glycolipids / Q000032:analysis*; D008163:Luminescent Measurements; D009195:Peroxidase
D006017:Glycolipids; D001710:Biotin; D009195:Peroxidase