beta-galactosidase assay using capillary electrophoresis laser-induced fluorescence detection and resorufin-beta-D-galactopyranoside as substrate.
beta-Galactosidase was incubated for 60 min with the fluorogenic substrate resorufin-beta-D-galactopyranoside, which is converted by the action of the enzyme into resorufin and galactose. A 160 pL aliquot of reaction mixture was analyzed by capillary electrophoresis utilizing laser-induced fluorescence detection. Based on the detection of the resorufin formed, the limit of detection of beta-galactosidase was 1.5 x 10(-15) M or 900 molecules of enzyme in a 1 microL sample.
Eggertson
M J
MJ
Department of Chemistry, University of Winnipeg, 515 Portage Avenue, Winnipeg, Manitoba, Canada R3B 2E9.
Craig
D B
DB
D019075:Electrophoresis, Capillary / Q000379:methods; D005697:Galactosides; D007834:Lasers; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D010078:Oxazines; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000032:analysis* / Q000378:metabolism
D005697:Galactosides; D010078:Oxazines; C056561:resorufin galactopyranoside; D001616:beta-Galactosidase