MALME-3M cells were incubated at 37°C for 24h with vehicle control (0 µM) and 1 µM andrographolide (ab120636). Increased expression of p53 in MALME-3M cells correlates with an increase in andrographolide 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 ab26 at 5 µg/ml and ab8227 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.