Hi Sir and Ma'am.

I rescently isolated a sample from probiotic strain and found that it suppress LPS induced TNF Alpha and IL-6 mRNA and proteuin secretion in RAW 264.7 cell.

But here is problem.

IL 1beta. Which is found to be significantly suppressed in qpcr, is found to be in congruent with the ELISA result.

As if secreted IL 1beta are degraded....

I have went though some areticles and found that IL 1beta protein level reaches max at 3~4 h of stimulation using LPS.

And a number of papers have used lesser stimulation time for IL 1beta for their ELISA experient. (Compared to other pro-inflammatory cytokines)

How long do you stimulate RAW 264.7 cells w/ LPS when preparing for IL-1beta AND TNF alpha?

In our lab, we usually stimulate LPS 24h for all types of cytokine

Best Regards.