Ethanol Fixation Protocol
Prepare 70 % Ethanol and chill to - 20°C ( NOTE : Do not freeze ethanol for long - term storage ) .
Prepare target cells of interest and wash 2X with PBS , centrifuging at 350xg for 5 minutes .
Discard supernatant and loosen the cell pellet by vortexing .
Add 3 ml cold 70 % ethanol drop by drop to the cell pellet while vortexing .
Continue vortexing for 30 seconds .
Incubate at - 20°C for 1 hour .
Wash 2X with BioLegend Cell Staining Buffer ( Cat . # 420201 ) and resuspend cells at 0.5 - 1.0 x 107 cells / ml .
Use 100 μl cell suspension / staining tube .
