Reagents

  • 2.5µg/ml Annexin V (FITC) [final concentration]
    • Binding Buffer
    • HEPES buffer: 10mM HEPES / NaOH, pH 7.4
    • 150mM NaCl
    • 5mM MgCl2
    • 5mM KCl
    • 1.8mM CaCl2
  • 50µg/ml PI (propidium iodide, Sigma Chemical cat# P-4170) in 1 x PBS

Staining

  1. Resuspend 5 x 105 cells in 500µl HEPES buffer.
  2. Add 2.5µg/ml Annexin V (FITC) [final concentration].
  3. Incubate at room temperature for 10 to 15 minutes.
  4. Add 10µl PI.
  5. Run on flow cytometer.
annexin_histo

 

 

 

 

 

 

Sample histogram using this protocol.