Differential Suppression of Background Mammalian Lysosomal Beta-Galactosidase Increases the Detection Sensitivity of LacZ-marked Leukemic Cells

Anal Biochem. 1994 Nov 1;222(2):456-60. doi: 10.1006/abio.1994.1516.

Abstract

A method is described for the detection of Escherichia coli beta-galactosidase-expressing leukemic cells in ex vivo bone marrow samples. 4-Methylumbelliferyl-beta-D-galactopyranoside is used as a substrate in a kinetic assay. D-Galactose is used to suppress endogenous lysosomal beta-galactosidase activity, yielding a sixfold increase in sensitivity. With this assay, the detection limit is one leukemic cell per 10(4) normal bone marrow cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Disease Models, Animal
  • Escherichia coli / enzymology
  • Galactose / chemistry
  • Galactosides / chemistry*
  • Genetic Markers
  • Hydrogen-Ion Concentration
  • Hymecromone / analogs & derivatives*
  • Hymecromone / chemistry
  • In Vitro Techniques
  • Kinetics
  • Leukemia, Myeloid, Acute / enzymology*
  • Leukemia, Myeloid, Acute / genetics
  • Leukemia, Myeloid, Acute / pathology
  • Lysosomes / enzymology*
  • Rats
  • Rats, Inbred BN
  • Sensitivity and Specificity
  • Substrate Specificity
  • beta-Galactosidase / analysis*
  • beta-Galactosidase / antagonists & inhibitors

Substances

  • Galactosides
  • Genetic Markers
  • Hymecromone
  • 4-methylumbelliferyl-galactopyranoside
  • beta-Galactosidase
  • Galactose