K562 cells were incubated at 37°C for 4h with vehicle control (0 μM) and 15 nM of neurokinin A (ab120185). Increased expression of p53 (ab26) correlates with an increase in neurokinin A 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 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.