Nuclei for Cell Staining Analysis
Hypotonic Staining Buffer:
|Sodium citrate tribasic dihydrate||10.0 mg|
|Triton® X-100||30.0 ul|
|Propidium iodide||5.0 mg|
|Ribonuclease A||0.2 mg|
- Aliquot 1x106 cells into each tube.
- Centrifuge 200 to 400 xg for 5 - 10 minutes and aspirate supernatant without disturbing the pellet.
- Add 500 ul of the Hypotonic Staining Buffer to the pellet and gently mix well.
- Keep samples at 4°C protected from light for 30 min or for a maximum of 1 h before acquisition on the flow cytometer.
Debris can increase with longer staining incubation times.
Krishan A: Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J. Cell Biol. 66:188-193, 1975.