I'm trying to measure the concentration of IL 1b in tracheal epithelial cells. Cells are grown in RPMI media. However, I have run the experiment several times and the amount of background is ruining the assay. I have tried spiking the RPMI media with the standards that came with the commercial kit (it is a pre-developed kit) and was expecting to see a concentration curve similar to the regular standards. However what was observed was a complete lack of detection. As you can see in the document the blue curve is the standards that were provided by the company, and the orange curve are the spiked standards. There is no gradual change in concentration when there is media present.

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