I am working on THP-1 cells. I subcultured THP-1 into 6-well plates with 3.5x10^5 cells/well. I used PMA 100ng/mL for 24h to activate THP-1 (M0), then changed the media to culture media without PMA or M1 media the following day. The supernatant will be harvested after 48h to measure TNFa by ELISA. Although TNFa in M1 condition is higher than that of M0 condition, in M0 (only treated with PMA), TNF-a is quite high, nearly 1000 pg/mL. My cells' condition is good and does not have any contamination. Does anyone have the same problems as me? And how to fix it>
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