I have started working with THP-1 cells.and have found different protocols regarding the duration of PMA treatment and then polarization to M1 and M2 macrophages. I got weird results with ELISA for TNF-alpha and IL-10. I have tried different concentrations of PMA from 20 nM to 200, and duration of 1 day to 3 days. Some people say that we have to add cytokines while PMA is present and some not. Has anyone a robust protocol?
Thanks in advance,