There are many recipes available. As long as they have sufficient calcium most of them should be suitable. We use the following: Binding buffer = 10 mM Hepes adjusted to pH 7.4 140 mM NaCl 2.5 mM CaCl2 Method: 1. Wash cells in PBS, centrifuge at 250g (10 min). 2. Resuspend cells in 192 ul of binding buffer. 3. Add 8 ul of Annexin V-FITC and incubate in the dark for 10 min. 4. Resuspend cells in 0.5 ml of binding buffer. 5. Add 10 ul of propidium iodide (10 ug/ml). 6. FACS within 1 hour.
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