A flow cytometric method for the rapid detection of beta-galactosidase transfected cells: an in vitro and in vivo study.
A flow cytometric method has been developed for the rapid analysis of lacZ transduced cells. The method described is based on an indirect immunofluorescence staining procedure using a monoclonal antibody which binds specifically to beta-galactosidase from E. coli and to beta-galactosidase fusion proteins. This technique was used for the quantification in vitro as well as in vivo of beta-galactosidase expression in B16 melanoma cells. The described method is appropriate for a variety of cell types (species, lineage), is simple, quantitative, reliable, rapid and applicable to all constructs containing the lacZ selectable markers. It should prove to be very helpful (1) for the quantification of cells expressing the lacZ reporter gene and (2) for studying gene regulation, including transfection modality, promoter efficacy, enhancer activity, and other regulatory factors.
Mouawad
R
R
Laboratory of the Medical Oncology Department, Salpetrière Hospital, Paris, France.
Khayat
D
D
Zerrouqi
A
A
Ghoumari
A M
AM
Soubrane
C
C
D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect; D014162:Transfection; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000032:analysis* / Q000235:genetics
D001426:Bacterial Proteins; D001616:beta-Galactosidase