Human/Mouse/Rat Phospho-ATM (S1981) Antibody

Detection of Human/Mouse/Rat Phospho-ATM (S1981) by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 1 µM camptothecin (CPT) for 1 hour. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human/Mouse/Rat Phospho-ATM (S1981) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1655) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-ATM (S1981) was detected at approximately 370 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. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Phospho-ATM (S1981) by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 1 hour, loaded at 0.2 mg/mL. A specific band was detected for Phospho-ATM (S1981) at approximately 270 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-ATM (S1981) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1655). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Detection of Human ATM by Western Blot

Resveratrol activates ATM in transformed human cell lines.(a, b) Human HEK293T cells were treated with 0.01 mM ATM inhibitor (KU-55933) or vehicle (DMSO) for 1 hour, followed by incubation in media also containing 0.1 mM resveratrol (or DMSO), as indicated. After 30 minutes incubation with resveratrol, H2O2 (0.1 mM) or bleomycin (1 µg/ml) was added as indicated for 30 minutes before harvesting. Western blotting was performed with with antibodies directed against ATM, phospho-ATM Ser-1981, p53, and phospho-p53 Ser-15 as indicated. (c, d) Human HEK293T cells were treated with resveratrol, bleomycin, or both as in (a) and probed for gamma-H2AX foci by immunofluorescence. Cell images (291, 267, 216, and 260 cells, respectively) were analyzed for foci using Image J software, and the average number of foci per cell as well as the percentage of cells containing >5 foci were quantitated. (e, f) Human HCT116 cells were incubated with KU-55933, resveratrol, H2O2, and bleomycin as in (a, b) but additional phosphorylation targets were examined using antibodies directed against Smc1, phospho-Smc1 Ser-957, Kap1, phospho-Kap1 Ser-824, Nbs1, phospho-Nbs1 Ser-343, Chk2, and phospho-Chk2 Thr-68 as indicated. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0097969), licensed under a CC-BY license. Not internally tested by R&D Systems.