I'm trying to produce cytokines(TNFalpha, IL-2, IL-4 IL-10, TGF-b and IFNgama) by stimulated PBMC. I isolated PBMC from whole blood by Ficoll method. In 6 well tissue culture plate, I added 10 ug/mL PHA_L (sigma) for 105cells/ml and incubated 3 days. I used cell culture superatants for ELISA assay but cytokine levels were very low. I have no experience and need advice.
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Cytokine production by stimulated PBMC
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