Fenofibrate, PPAR-alpha agonist (ab120832)

HL-60 cells were incubated at 37°C for 24h with vehicle control (0 µM) and different concentrations of fenofibrate (ab120832). Increased expression of ACADM in HL-60 cells correlates with an increase in fenofibrate concentration, as described in literature.

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 µg 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 5% BSA before being incubated with ab110296 at 1 µg/ml 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.