Which is the best time point to measure the expression of CD56, CCR7, CD45RA, FOXP3, CD69, CD25 on human T cells?
Even though your question is very clear -cut. but the cell-culture could be produce and analyzed with 6-10 days in most of the cases mentioned.
It all depends on what type of stimulant you are using for your experiments.
The problem I think is that some of them are expressed early and others lately and I would like to find a unique time point in which I can meaure them in which they are "good" expressed to do an antibody titration for FACS analysis
Hello Lorenzo, I assume you mean characterisation after activation rather than steady state evaluation, which I would want to perform fresh.Generally, 5-7 days incubation prior to Elispot, or flow cytometry evaluation, is the period reported to evaluate an activation of phenotype following antigen stimulation, though elevation of CD 25 and CD69 can be seen earlier. Mitogen activation usually maximal by 2-3 days depending on agent used, PHA, ConA, etc. For a long term study I would consider doing a time course assessment fresh, d1, .... dX to see where there were expression/intensity plateaus for the antigens to be evaluated.
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