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A rapid, quantitative and inexpensive method for detecting apoptosis by flow cytometry in transiently transfected cells
Author(s) -
Gábor M. Lamm
Publication year - 1997
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/25.23.4855
Subject(s) - propidium iodide , biology , transfection , flow cytometry , microbiology and biotechnology , green fluorescent protein , apoptosis , cytometry , population , plasmid , cell , dna , gene , programmed cell death , genetics , sociology , demography
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.

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