Hi,
I have fixed with 4% PFA Raji cells, and stored at -80 C in 10% DMSO + 90% HSA. I would like to do a titration of the CD19 antibody to see if the cells express this marker. I am writing my experimental plan , but cannot find any protocol that i can base my experiment on or any literature.
any tips on how I can do this?
Thank you!
Fixation prior to staining the cells with your CD19 antibody possibly could alter the epitope the antibody binds and therefore you would not see a positive signal by flow cytometry.
You could thaw the cells and try staining them/running flow but if it doesn't work then what I described is probably the cause of a negative result. Be sure to include a positive control for CD19 so you know that the antibody works/is not the cause of a negative signal.
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