Human Phospho-p53 (S15) Antibody

Detection of Human Phospho-p53 (S15) by Western Blot.

Western blot shows p53 immunoprecipitated from lysates of MCF-7 human breast cancer cell line using Human/Mouse/Rat p53 Agarose-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # GAF1355). MCF-7 cell line was untreated (-) or exposed (+) to 10 Gy ionizing radiation (IR) for 3 hours. PVDF membrane was probed with 1-2 µg/mL Mouse Anti-Human Phospho-p53 (S15) Monoclonal Antibody (Catalog # MAB1839) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for Phospho-p53 (S15) was detected at approximately 53 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U ?-phosphatase (?-PPase) for 1 hour. For additional reference the membrane was stripped and reprobed with 1:5000 dilution Human/Mouse/Rat p53 HRP-conjugated Antigen Affinity-purified Polyclonal Antibody HAF1355 (lower panel, Catalog # HAF1355) This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of p53 in camptothecin-treated MCF‑7 Human Cell Line by Flow Cytometry.

MCF-7 human breast cancer cell line was unstimulated (light orange open histogram) or treated with 1 µM camphtothecin for 6 hours (dark orange filled histogram), then stained with Mouse Anti-Human Phospho-p53 (S15) Monoclonal Antibody (Catalog # MAB1839) or isotype control (Catalog # MAB002, blue open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.