Abstract
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.
MeSH terms
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Adamantane / analogs & derivatives
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Cell Line
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Colorimetry / methods
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Escherichia coli / genetics
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Galactosides
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Genes, Bacterial
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Genes, myc
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Genetic Vectors
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Humans
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Hydrogen-Ion Concentration
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Indicators and Reagents
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Infant, Newborn
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Kinetics
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Luminescent Measurements
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Lysosomes / enzymology
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Microchemistry / methods
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Promoter Regions, Genetic
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Substrate Specificity
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Transfection*
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beta-Galactosidase / analysis*
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beta-Galactosidase / genetics*
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beta-Galactosidase / metabolism
Substances
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Galactosides
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Indicators and Reagents
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3-(4-methoxyspiro(1,2-dioxetane-3,2'-tricyclo(3.3.1.1(3,7))decan)-4-yl)phenylgalactopyranoside
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beta-Galactosidase
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Adamantane