4.4. Measurement of 12(S)-HETE Level

The MDCK cells were incubated with the indicated concentration of IS. The cellular content of 12-hydroxyeicosatetraenoic acid (12(S)-HETE) was determined using a commercial quantitative immunoassay kit (Biomerica, Newport Beach, CA, USA) according to the manufacturer’s protocol. Homogenized cell extracts, as prepared for Western blotting, were diluted with buffer diluents supplied in the kit, as previously described [16]. The total amount of protein in the sample was quantified using the ELISA immunoassay method. The amount of 12(S)-HETE in the cell is expressed as μg per g of protein.