I'm interested in staining cell's dry mass and measuring it on flow cytometry, nevertheless I'd like to know which is a suitable time for the CFSE's intake to the cells. Other suggestions for measuring dry mass are accepted.
Greetings
Abcam suggests to stain live cells for only 10 min with CFSE
http://www.abcam.com/ps/products/113/ab113853/documents/ab113853%20CFSE%20Cell%20Labeling%20Kit%20V2%20(website).pdf
CellTrace - suggests to stain for 20 min, but it also depends on cell number and dye concentration (at room temperature or 37°C),
https://tools.thermofisher.com/content/sfs/manuals/celltrace_cell_proliferation_kits_man.pdf
I was staining 1 million MCF-7 cells for 30 min at 37C and it also worked well. You need to wash cells to get rid of unloaded dye.
Dear Steven, please see the attached protocol for CFSE staining using CellTrace. I've gotten it from a neighboring lab and it ALWAYS worked fabulously for me. It takes you through all the necessary steps, but the optimization is for primary B cells. (In fact I have used it for all kinds of cells, but never changed the staining procedure itself). You can give it a try with HEK293 and let me know how it works.
All the best and good luck with your experiment!
Mika
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