1. Harvest cell using 0.25% trypsin
2. Resuspend in 1ml DMEM/F-12 media with Trypsin inhibitor
3. Placed in a 1.5 ml micro centrifuge tube and spin down
4. Wash with FACS buffer (1XPBS, 5% FBS, and 0.1% sodium azide) in ice
5. Resuspend in 50-100 μl and incubate in ice or 4°C for 30 min ~ 1 hour with appropriate antibody (1:20-1:40 dilution)
6. Wash three times with FACS buffer
7. Incubate with a flourescein-conjugated IgG secondary antibody(1:100) for 30 min.
8. Wash three times with FACS buffer
9. Add 0.5 ~ 1 ml of cold 1% Paraformaldehyde solution to the pellet.
10. Suspense cell immediately
11. Store the fixed cells at 4°C in the dark
12. These cells may be stored for at least one week prior to FACS analysis
IMPORTANT: Protect from direct light