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

Anal Biochem. 1991 Nov 15;199(1):119-24. doi: 10.1016/0003-2697(91)90278-2.

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.

Publication types

  • Comparative Study

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*
  • beta-Galactosidase / genetics*
  • beta-Galactosidase / metabolism

Substances

  • 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