Propidium Iodide Staining for FACS
Propidium Iodide Staining for Fluorescence-Activated Cell Sorting (FACS)
- Collect 2x106 cells.
- Resuspend cell pellet in 2 ml PBS.
- Vortex gently while adding 5 ml of 95% ethanol.
- Fix at least 30 minutes at RT.
- Store at 4°C (or -20°C) until ready to stain.
Staining
- Spin cells out of fix (3000x 5 minutes).
- Resuspend pellet in 1 ml of PI Working Solution.
- Add 100 µl of 1 mg/ml DNase-free Rnase.
- Incubate at 37°C for 30 minutes.
- Samples are ready for flow cytometry (FCM) analysis. They may be stored in refrigerator O/N.
Solutions
Note: For a single FCM run, use the same PI working solution to reduce variation between samples.
Stock Propidium Iodide (PI)
- 500 µg/ml made in H20
Working Solution
- 1:10 dilution with PBS or Tris buffer