Intracellular staining for FACS

(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