Instructions For UseImportant:
Thaw all components prior to use.Note:
The following protocol is a guideline and it should be modified for each experiment as needed.
1. Prepare a 200 μM stock solution by adding 892.5 μl of DMSO to a CFDA-SE vial and mix by vortexing.
2. Create a working solution (5-0.5 μM) by diluting the CFDA-SE stock solution from step 1 with your buffer of choice at pH 7.
3. Resuspend 1 x 106
cells of interest in 500 μl of the working solution.
4. Incubate the cells for 5-20 min at room temperature. Protect from light.
5. Centrifuge the sample and remove the supernatant.
6. Wash the pellet with 3 ml of your buffer of choice.
7. Resuspend the cells in 500 μl of fresh, prewarmed culture media.
8. Remove 200 μl to analyze for time zero.
9. Place the remaining cells in the appropriate conditions for cell proliferation.
10. Harvest the cells and stain them for the other markers if desired.
11. Analyze or sort the cells using a flow cytometer or S3™ cell sorter
with a 488 nm laser.