I am looking for a standard protocol to stimulate THP-1 cells by immune complexes (IC). My aim is to demonstrate Interleukin-1 beta secretion from IC-stimulated THP-1 cells by sandwich ELISA.
Hi!
Are you going to differentiate those cells to macrophages?
You can stimulate IL-1beta secretion with LPS.
You'll have to do a time-concentration response to see in which LPS concentration your cells are going to produce this cytokine.
Good luck!
Thanks for the suggestions. LPS can be used as a positive control for my study. But my aim is to show that the immune complex which I study can stimulate THP-1 cells to secrete pro-inflammatory cytokines. In PMA-differentiated THP-1 cells, I don't see any difference in TNF-alpha values between the test wells where immune complex is added and the control wells. Presently I am trying to immobilize the immune complex in 96 well plate and add THP-1 cells into it. In my preliminary studies I can see that TNF-alpha is released from THP-1 cells added to immobilized immune complexes. But there is no release of Interleukin-1 as determined by ELISA. I do ELISA after 24 hour culture.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology