Measuring Apoptosis using Annexin V and 7AAD

Cells can be stained for early stage apoptosis using Annexin V and late stage apoptosis using 7AAD or PI. Dyes can be combined further to differentiate these cells. Our in-house protocol uses Annexin V -FITC and 7AAD, however your assay procedure may use a different scheme.

  1. Resuspend cells in cold Annexin V Binding buf fer at a final concentration of 1e5 to 1e7 cells/mL.
  2. Transfer 100µl of cells to FACS tubes (12x75mm tubes).
  3. Add 5µl of FITC Annexin V.
  4. Add 5µl of 7-AAD viability staining solution.
  5. Gently vortex the tubes and incubate for 15min at room temperature in the dark.
  6. After incubation, add 400µl of Annexin V Binding buffer to each tube.
  7. Send samples to AVIVA for analysis by flow cytometry.