A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells.
An optimized chemiluminescent assay for beta-galactosidase using a chemiluminescent substrate AMPGD (3-(4-methoxyspiro[1,2-dioxetane-3,2'-tricyclo-[3.3.1. 1(3,7)]decan]-4- yl)phenyl-beta-D-galactopyranoside) is described. This assay is rapid and sensitive and can detect as little as 2 fg of beta-galactosidase. Its use for the quantitation of beta-galactosidase in cells transfected with lacZ-expressing vectors is described. It is possible to detect a single cell stably expressing lacZ by this technique.
Jain
V K
VK
Pediatric Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Magrath
I T
IT
D000218:Adamantane / Q000031:analogs & derivatives; D002460:Cell Line; D003124:Colorimetry / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005697:Galactosides; D005798:Genes, Bacterial; D016259:Genes, myc; D005822:Genetic Vectors; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007231:Infant, Newborn; D007700:Kinetics; D008163:Luminescent Measurements; D008247:Lysosomes / Q000201:enzymology; D008832:Microchemistry / Q000379:methods; D011401:Promoter Regions, Genetic; D013379:Substrate Specificity; D014162:Transfection; D001616:beta-Galactosidase / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism
D005697:Galactosides; D007202:Indicators and Reagents; C073981:3-(4-methoxyspiro(1,2-dioxetane-3,2'-tricyclo(3.3.1.1(3,7))decan)-4-yl)phenylgalactopyranoside; D001616:beta-Galactosidase; D000218:Adamantane