Flow Cytometry Immune Monitoring Shared Resource


Apoptosis with Annexin/PI

Binding Buffer:

  • 10mM HEPES/NaOH, pH 7.4
  • 150mM NaCl
  • 5mM KCl
  • 1mM MgCl2
  • 1.8mM CaCl2

PI Solution:

  • Propidium iodide 100µg/ml in Binding Buffer
  1. 1Wash 2x106 cells with PBS.
  2. Dilute FITC-Annexin V to 1µg/ml in binding buffer and resuspend cells in 1ml of this solution (prepared fresh each time)
  3. Incubate 10 min. in the dark at room temperature
  4. Add 10µl of PI solution for a final concentration of 1µg/ml prior to analysis