FACS Analysis

 

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 4C 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 4C in the dark

 

12. These cells may be stored for at least one week prior to FACS analysis

 

IMPORTANT: Protect from direct light