Hello everyone,
I want to do an ELISA assay to detect IFN gamma and IL2 secreted from a primary T cell line.
In order to do so, on day 1 I will thaw these T cells and centrifuge them to get rid of the DMSO and then resuspend them in media consisting of : RPMI, FBS 10%, PenniStrep 1%, 0.5% non essentials amino acids, 1% HEPES, 0.5% L-Glutamine and 0.001% B-mercaptoethanol and 2nm of IL2.
Then on day 2 I will put those cells in 96 well plates with the APC that have been incubating with the antigen prior in the same media.
This incubation will last 48h at 37°C and then I will harvest the supernatant after centrifugation of the plate to perform an ELISA assay.
I read that feeder cells are important when working with primary cells but I don't know if I have to add them when working with such short time, if I don't put any will it prevent the T cells to activate and secrete IL2 and IFN gamma ?
Thanks in advance, I hope I was clear enough.
Best regards,
Sonia
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