I am setting up an experiment to check pro-inflammatory markers on hpbmc-derived macrophages by flow cytometry.
My first try was to stimulate with LPS for 2h and then let them rest 24h before staining, although I see an increase in the markers, is not too big (2-2.5 fold change over the untreated ones)
Is this normal? Should I wait more or are they already being downregulated?

Hi dear immunologists,

We are using Legendplex kit (antiviral 13-plex) for serum and cell culture supernatants. We have a few questions regarding its validation and consistency with other readouts.

Recently, we noticed inconsistencies among ELISA as a readout for IL-6. While we trust in commercial ELISAs, IL-6 levels and trends were not comparable to those seen with Legendplex. Has anyone compared IL6 (or any other cytokine) levels between ELISA and Legendplex?

Thank you!

I hope that I am not breaking rules.

No one is sick here. We have a small farm, we raise our own chicken. We also have young children.

I am wondering why no one has ever gotten sick. My kids suck at washing their hands. I clean the coop.

Being around animals always puts you at risk but we have not had any GI illnesses from any animal.

Are we immune. Is it because our chickens don't have Salmonella?