Sample experiment using ab128570 on MCF-7 cells treated with for 6 hours with Camptothecin or drug vehicle (DMSO). The data (presented as mean +/- SEM) were analyzed as described in Data Analysis section.
Specificity of antibodies demonstrated by immuno-cytochemistry. Primary antibodies used in this assay kit were validated by staining MCF-7 cells treated with Camptothecin or vehicle and imaged by fluorescent microscopy. An increase in both p53 phospho S46 and total p53 protein signals upon Camptothecin treatment.
Antibody specificity demonstrated by Western Blot Analysis. Primary antibodies used in this assay kit were validated by Western Blotting using Hek293T induced with Etoposide (to increase the levels of p53 phosphorylation). Cell extract was treated with λ protein phosphatase (lane 2) or left untreated (lane 1). Samples were analyzed by Western blotting using the rabbit p53 phospho S46 antibody (A). The membrane was re-probed with the mouse total p53 protein antibody (B). Note the sensitivity of the p53 phospho S46 signal to treatment with λ protein phosphatase and resistance of the total p53 protein signal to this treatment.