7-Aminoactinomycin D for apoptosis staining in flow cytometry.
7-Aminoactinomycin D (7-AAD) is a DNA dye that distinguishes viable, apoptotic, and late apoptotic/dead cells in flow cytometry. Several staining protocols using 7-AAD have been described, but data on the influence of the 7-AAD concentration on the readout are not available. Therefore, we compared the results obtained by staining with 1, 5, 10, and 20μg/ml 7-AAD for 20min with the PE-Annexin V Apoptosis Detection Kit and Cell Death Detection ELISA(PLUS) in lymphocytes and CEM human leukemia cells. The results showed that 7-AAD staining with 5, 10, and 20μg/ml, but not with 1μg/ml, is suitable for quantification of apoptosis in flow cytometry.
Department of Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, 69120 Heidelberg, Germany.
Nadine C L
NC
Zembruski
Vanessa
V
Stache
Walter E
WE
Haefeli
Johanna
J
Weiss
D017209:Apoptosis; D003609:Dactinomycin / Q000031:analogs & derivatives* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D003609:Dactinomycin; C025942:7-aminoactinomycin D