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[KR1972]

Hi all;

I am new (both Forum and cytokines!!).

We are trying to detect cytokines from cultured/stimulated PBMC.
We are incubating in triplicate for 5 days.

Do you pool your supernatants or Not?

Currently, we are not pooling the supernatant and running each individual well with a CBA and we are observing quite variations between wells.
We only run them once.

So what are your best practices?

Pooled supernatants and run in duplicates?
Unpooled supernatants and run once?


Thank you very much for your help!!!!