7-Aminoactinomycin D for apoptosis staining in flow cytometry

Anal Biochem. 2012 Oct 1;429(1):79-81. doi: 10.1016/j.ab.2012.07.005. Epub 2012 Jul 14.

Abstract

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.

MeSH terms

  • Apoptosis*
  • Dactinomycin / analogs & derivatives*
  • Dactinomycin / metabolism
  • Flow Cytometry / methods*
  • Fluorescent Dyes / metabolism*
  • Humans
  • Lymphocytes / cytology
  • Lymphocytes / metabolism
  • Staining and Labeling / methods*

Substances

  • Fluorescent Dyes
  • Dactinomycin
  • 7-aminoactinomycin D