HepG2 cells were incubated at 37°C for 24h with vehicle control (0 μM) and different concentrations of clotrimazole (ab120744) in DMSO. Increased expression of MRP3 (ab3375) correlates with an increase in clotrimazole concentration, as described in literature.

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab3375 at 1/50 dilution and ab9484 at 1 μg /ml overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (ab97040) at 1/10000 dilution and visualised using ECL development solution.