|Detection of Human Phospho-p53 (S37) 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 treated (+) with 1 μM camptothecin (CPT) for 5 hours. PVDF membrane was probed with 0.4 µg/mL Rabbit Anti-Human Phospho-p53 (S37) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3306) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-p53 (S37) was detected at approximately 53 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda -phosphatase ( lambda -PPase) for 1 hour. For additional reference the membrane was stripped and reprobed with 1:5000 dilution of Human/Mouse/Rat p53 HRP-conjugated Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # HAF1355) This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
The p53 tumor suppressor protein acts to enforce cell cycle checkpoints or signal apoptosis in cells that have incurred genotoxic damage. The ATR and DNA-PKCS kinases phosphorylate p53 at serine 37 in response to UV light and other genotoxic stressors. This phosphorylation event results in increased p53 transcriptional activation.
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