Apoptosis with AnnexinV/PI

 

Binding Buffer:

10mM HEPES/NaOH, pH 7.4

150mM NaCl

5mM KCl

1mM MgCl2

1.8 mM CaCl2

 

PI solution:

Propidium iodide 100µg/ml in Binding Buffer

 

Wash 2x106 cells with PBS.

Dilute FITC-Annexin V to 1 µg/ml in binding buffer and resuspend cells in 1 ml of this solution (prepared fresh each time).

Incubate 10 min. in the dark at room temperature.

Add 10 µl of PI solution for a final concentration of 1 µg/ml prior to analysis.