- PBS or Hanks (Ca/Mg ++ free)
- 25mM Hepes pH= 7.0
- 0.5-2% FBS (heat inactivated)
- 1mM EDTA.
- 0.2 mm sterile filtered
Culture Media is not good as a sort buffer because outside the confines of the incubator, the pH tends to become basic, which may be the reason for low viability and recovery. The addition of Hepes will help to stabilize the pH.
For cells that don’t stick together, you can modify the sort buffer to omit the EDTA. You should use the least amount of FBS the cells need to remain happy.
The addition of EDTA will help reduce the stickiness of some cell types. The concentration of EDTA should not exceed 5mM.
Place on ice or store at 4C until use. You can make up 1 L at a time and store at 4C, as long as it is kept sterile for staining cells.
To make 1L:
2ml 0.5mM EDTA
0.1% sodium azide
1x PBS to 1L
Filter and sterilize if needed.