Fluo-3 and Fluo-4 Protocols
1. Pellet cells and adjust to 10 x 106/ml.
2. Dissolve 50 mg freeze-dried
aliquot of Fluo-3 (Invitrogen, Cat. No. F1242) with
100 ml DMSO.
3. Or dissolve 50 mg
freeze-dried aliquot of Fluo-4 (Invitrogen, Cat. No.
F14201) with 100 ml DMSO.
4. Incubate lymphocytes with 1-2 mM Fluo-3/4, granulocytes 2-3 mM, dendritic
cells at 4 mM at 370C
for 45 min.
5. Wash twice in calcium free PBS.
6. Adjust cells to 1 x 106/ml in Falcon 5ml tubes.
7. Run cell samples with 530 nm channel in linear mode against
time.