Example positive control sample standard curve. A dilution series of extract in Incubation Buffer in the working range of the assay. The extract was prepared from Hek293T cells treated with etoposide by direct lysis.
Example experimental analysis of drug treatment of Hek 293T and MCF7 cells. Cells were treated with etoposide, camptothecin or drug’s vehicle, as indicated. Diluted cell extracts were analyzed by this kit (A, C) and p53 protein ELISA (B, D), using ab117995. Extract of Hek 293T treated with etoposide was used for positive control sample standard curves. Relative levels interpolated from standard curves are shown.
The p53 Phospho S6 ELISA specifically measures the phosphorylated Serine. Extracts of Hek 293T cells (induced with etoposide) were treated with increasing concentrations of λ protein phosphatase (400x= 400-times diluted, 100x=100-times diluted), mock-treated (Mock) or left untreated (Contr), and phospho Ser6 (in red) and total p53 protein (in green) levels were determined, respectively, using this kit and ab117995. Dilutions of extracts of Hek 293T cells treated with etoposide were used to construct the standard curves.
The detector antibody used in this kit specifically detects the phosphorylated p53 as determined by Western blotting. Extracts of MCF7 cells (20 µg/lane) were treated with vehicle (lane 2) or Camptothecin (lane 3). Extracts of Hek293T cells (20 µg/lane) were treated with vehicle (lane 4) or Etoposide (lane 5). Extracts of Hek 293T cells (induced with Etoposide, 8 µg/lane) were treated with increasing concentrations of λ protein phosphatase (lane 7, 400x diluted; lane 8, 100x diluted; lane 9, 25x diluted) or left untreated (lane 6).
The capture antibody used in this kit specifically detects the phosphorylated p53 as determined by Western blotting. Extracts of MCF7 cells (20 µg/lane) were treated with vehicle (lane 2) or Camptothecin (lane 3). Extracts of Hek293T cells (20 µg /lane) were treated with vehicle (lane 4) or Etoposide (lane 5). Extracts of Hek 293T cells (induced with etoposide, 8 µg /lane) were treated with increasing concentrations of λ protein phosphatase (lane 7, 400x diluted; lane 8, 100x diluted; lane 9, 25x diluted) or left untreated (lane 6).