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NUCLEI FOR CELL CYCLE 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
|
| H20 |
100
ml
|
- 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.
Note: Debris can incease
with longer staining incubation times.
Reference: Krishan
A: Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium
iodide staining. J. Cell Biol. 66:188-193, 1975.
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