Flow Cytometry Laboratory

Annexin V FITC Staining protocol

1. Take cell suspension and spin.

2. Resuspend in 1ml of Hepes buffer, spin again.

3. Resuspend in 1ml Hepes buffer and add 2.5 g/ml of Annexin V-FITC.

4. Incubate in the dark at room temperature for 30 minutes.

5. Analyse after adding Propidium Iodide to gate out dead cells.

This method is only really applicable to suspension cells and should identify intermediate apoptotic cells.

Hepes Buffer

10 mM Hepes/NaOH pH 7.4 476.6 mg/200ml

140 mM NaCl 1.64 g/200ml

2.5.mM CaCl2 73.5 mg/200ml

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