A rapid, quantitative and inexpensive method for detecting apoptosis by flow cytometry in transiently transfected cells.
We describe a rapid and quantitative flow cytometric method for determining the apoptotic or anti-apoptotic potential of a gene in various cell types. A plasmid carrying green fluorescent protein (GFP) is co-transfected with an expression vector encoding the gene of interest. Subsequently cells are stained with propidium iodide and, utilising flow cytometry, transfected, GFP-expressing single cells are detected and apoptotic cells in this population are identified by their DNA content of <2 N. The method detects apoptosis as reliably as established methods using in situ nick-end labelling but is faster, easier and less expensive.
Lamm
G M
GM
Research Institute of Molecular Pathology (I. M. P.), Dr. Bohr-Gasse 7, A-1030 Vienna, Austria.
Steinlein
P
P
Cotten
M
M
Christofori
G
G
D000818:Animals; D017209:Apoptosis; D005434:Flow Cytometry / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000737:chemistry; D014162:Transfection; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins