MCF7 cells were incubated at 37°C for 6h with vehicle control (0 µM) and different concentrations of honokiol(ab120647). Increased expression of acetyl coenzyme A carboxylase (phospho S79)(ab31931) in MCF7 cells correlates with an increase in honokiol 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 5% BSA before being incubated with ab31931 at 1 µg/ml and ab8227 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.
MCF7 cells were incubated at 37°C for 6h with vehicle control (0 µM) and different concentrations of honokiol(ab120647). Increased expression of AMPKα 1 + AMPKα 2 (phospho T172) (ab23875) in MCF7 cells correlates with an increase in honokiol 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 5% BSA before being incubated with ab23875 at 1/1000 dilution and ab8227 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.