if I have THP-1 cells what concentration of LPS need to use for induce the cells and get NLRP3 inflammasome? some article says 10ng/ml for 1*106 cells/ ml is this correct
To stimulate PMA-treated THP-1 cells with LPS, I would use 100 ng/ml for 3-4 h (see our paper in the link below).
Link: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0027040
I have also used 100 ng/mL overnight, followed by washing the following day. Be aware, though, that PMA concentration can effect cellular responses and may need to be optimized for your particular assay. This paper may be of some interest: http://link.springer.com/article/10.1007/s00011-007-6115-5#page-1
The cited paper by Dr. Yaron could be helpful.
We prime the THP-1 with 5 ng/mL PMA for 5-7 days to yield differentiated cells. They become very large and adherent, though fractions of them will die off during differentiation.
I agree with Dr. Choubey that LPS of that range should be enough. You can consult the inflammasome literature easily in Cell or Nature series papers.
Try to prepare the working conc. of the LPS freshly.
100 ng/mL for 4-6 hours provides the optimal response for pro-IL-1B and NLRP-3 expression
may I ask as well, which are the best concetrations of LPS and ATP to induce THP-1 cells without PMA treatment in inflammasome activation assay? also, how many cells should I use? some papers say 500000, others less, others more...many thanks
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