Does anybody know what happens to TNF alpha in PBMC after LPS stimulation? Or in MLR assay after LPS stimulation? Over more than 24 h i.e. up to 7 days?
my experience with TNFa secretion is that it is sometimes secreted in a biphasic manner. you'll probably see a rapid increase in secretion at 4-6 hours thatl'' gradually decline.
If your assay is to run for 7 days, will you be changing the media during that time? if so, will LPS be added with each media change?
If you are not changing the media, or are not going to re-dose with LPS, you will probably not see any TNF secretion beyond the first day or so.
If you add LPS to the cells just once, you can expect a peak in TNF-a protein levels within the 1st 24 h and TNF secretion by the cells will decline thereafter. By day 2-3 the increased secretion will be decline to baseline.
Thanks everyone. @caroline, would you not detect any tnf in the culture sups after 7 days? Is tnf consumed? Thanks for the help
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