MEF1 cells were incubated at 37°C for 24h with vehicle control (0 µM) and 5 µM of glibenclamide (ab120267) in DMSO. Increased expression of JNK1+JNK2 (phospho T183 + Y185) (ab4821) correlates with an increase in glibenclamide concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg 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 ab4821at 1/1000 dilution and ab85139 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.