- Liberate cells from tissue culture flask by trypsin digestion and collect in a 15 ml conical tube.
- Collect cells by centrifugation 3000xg 4 min and resuspend in 1 ml PBS.
- Repeat PBS wash.
- Add 3 ml 70% ethanol (-20oC) slowly DROP-WISE directly from the freezer while vortexing VERY GENTLY (No. 2 vortex setting on mixer).
- Incubate on ice for 30 min
- Collect cells by centrifugation 3000xg 2 min and resuspend in 1 ml Flow Buffer or PBS.
- Adjust volume to adjust the cell concentration.
- Store tightly capped at 4oC.