For monocyte stimulation it is mandatory to use materials that do not contain assayable levels of endotoxin to ensure that interactions with the TLRs and subsequent release of e.g. IL-6 is due to a contaminant in the test sample only. The assay should be prepared with materials that do not contain assayable levels of endotoxin which means that endotoxin contents are below the detection limit of the MAT assay.

In addition, the cell culture step must be prepared aseptically using sterile materials and environment, e.g. a laminar flow hodd.

Once the supernatant from pMAT cell stimulation is harvested there is no further need for aseptic or endotoxin-free handling. The ELISA can be conduceted on a lab bench.