I seeded 1X105 cells in 96 well plates in RPMI 1640+ 10% FBS medium and also with my different stimulus, although I got difference between groups but I saw that even unstimulated cells produce lot of IL-12p40 and TNF alpha but lower levels of IFN gamma after 24 hrs of incubation
What stimulation have you used? antigenic stimulation or cell specific stimulation. In my knowledge IFN-g is mainly produced by activated T cells. IL-12 is produced to LPS stimulation whereas IFN-g is produced to PHA or PMA or other antigens.
Hi my concern is I got IL12p40 and TNF alpha from the culture supernatant of unstimulated cells! Why is that?
I am not getting it clear. Whether you are getting higher IL-12 and TNF-alpha in unstimulated compared to stimulated ?
No, but the baseline level of IL12 p40 in unstimulated PBMC is higher but not more than stimulated.
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