A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells.

Analytical biochemistry ·Vol. 199 ·No. 1 ·1991-11-15 ·Pages 119-24

Jain VK, Magrath IT

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

Adamantane/analogs & derivatives Cell Line Colorimetry/methods Escherichia coli/genetics Galactosides Genes, Bacterial Genes, myc Genetic Vectors Humans Hydrogen-Ion Concentration Indicators and Reagents Infant, Newborn Kinetics Luminescent Measurements Lysosomes/enzymology Microchemistry/methods Promoter Regions, Genetic Substrate Specificity Transfection beta-Galactosidase/analysis,genetics,metabolism

Chemicals

Galactosides Indicators and Reagents 3-(4-methoxyspiro(1,2-dioxetane-3,2'-tricyclo(3.3.1.1(3,7))decan)-4-yl)phenylgalactopyranoside beta-Galactosidase Adamantane

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Jain V K

Pediatric Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.

Magrath I T

Article Info

Journal

Analytical biochemistry

Abbr.

Anal Biochem

ISSN

0003-2697

DOI

10.1016/0003-2697(91)90278-2

Published

1991-11-15

Pages

119-24

Language

English

Region

United States

NLM ID

0370535

Subset

External Links

PubMed source DOI: 10.1016/0003-2697(91)90278-2 View journal details

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A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells.

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