Rhod-2 Protocol

1. Pellet cells and adjust to 10 x 10⁶/ml.

2. Dissolve 50 mg freeze-dried aliquot of Rhod-2 (Invitrogen, Cat. No. R1245MP) with 50 ml DMSO.

1. Incubate cells with 0.5 mg/ml Rhod-2 at 37⁰C for 45 min.

2. Wash twice in calcium free PBS.

3. Adjust cells to 1 x 10⁶/ml in Falcon 5ml tubes.

4. Run flow cytometer with 575 nm channel on linear against the Time parameter.