(done on i.29 and primary B, MMCLs -NP-)
- pellet cells
- Fix cells 10' in Formaldehyde 3.7% (in PBS)
- add excess PBS and pellet cells 1800 rpm 5' (increased rpm to recover all cells after fixation)
- Permeabilize 10' in 0.1% Triton X-100
- Pellet cells 1800 rpm 5'
- Block in 1% goat serum in PBS 15'
- add 1ry Ab RT
- pellet cell 1800 rpm 5'
- add 2ry Ab 15' RT
- wash and resuspend in PBS, go to FACS