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 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