Anti-phospho-ATM (Ser1981) Antibody, clone 10H11.E12, Ascites Free

350.7 kDa calculated.

Serine-protein kinase ATM (EC 2.7.11.1; UniProt Q13315; also known as A-T mutated, Ataxia telangiectasia mutated) is encoded by the ATM (also known as AT) gene (Gene ID 472) in human. Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are related kinases that regulate cell cycle checkpoints and DNA repair. Mutation in the ATM gene results in the autosomal recessive disease ataxia telangiectasia (AT). Known ATM substrates include p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1, and Chk1. The S/TQ sequence constitutes the essential substrates phosphorylation site motif. Hydrophobic amino acids at positions -3 and -1, and negatively charged amino acids at position +1 are positive determinants for substrate phosphorylation, while positively charged residues surrounding the S/TQ are negative determinants. The complex phenotypes of cells derived from patients with AT suggests that ATM has additional cellular substrates. ATM is present as an inactive homodimer or multimer prior to activation. DNA double-stranded breaks induced by ionizing radiation (IR) cause rapid ATM autophosphorylation at Ser1981 in human or the Ser1987 equivalent in mouse.

Epitope: pSer1981.

Synthetic peptide corresponding to a.a. 1974-1988 of human ATM with phosphorylated Ser1981 (SLAFEEG[pS]QSTTISS).

Anti-phospho-ATM (Ser1981) Antibody, clone 10H11.E12, Ascites Free is an antibody against phospho-ATM for use in Immunocytochemistry, Immunoprecipitation, ELISA, Western Blotting.

Immunoprecipitation Analysis: A representative lot detected Ser1981-phosphorylated ATM interaction with endogenous EphA5 by co-immunoprecipitation using chromatin-enriched protein fractions of irradiated NCI-H460 human lung cancer cells (Staquicini, F.I., et al. (2015). J. Biol. Chem. 290(12):7345-7359).
ELISA Analysis: a representative lot was employed as the capture antibody for the detection of interaction between EphA5 and Ser1981-phosphorylated ATM. ATM pSer1981 captured from NCI-H460 human lung cancer cell extracts interacted with recombinant EphA5 cytoplasmic domain, but not with EphA5 extracellular domain or kinase domain (Staquicini, F.I., et al. (2015). J. Biol. Chem. 290(12):7345-7359).
Immunocytochemistry Analysis: A representative lot detected radiation-/IR-induced upregulation of nuclear ATM pSer1981 foci by fluorescent immunocytochemistry staining of methanol-fixed NCI-H460 human lung cancer cells (Staquicini, F.I., et al. (2015). J. Biol. Chem. 290(12):7345-7359).
Immunocytochemistry Analysis: A representative lot detected radiation-/IR-induced ATM Ser1981 phosphorylation by fluorescent immunocytochemistry staining of 1.2% formaldehyde-fixed, 0.1% Triton X-100-permeabilized HT29 cells (Bardelle, C., and Boros, J. (2012). J. Biomol. Screen. 17(7):912-920).
Immunocytochemistry Analysis: A representative lot detected ionizing radiation-/IR-induced ATM Ser1981 phosphorylation by fluorescent immunocytochemistry staining of methanol/acetone-fixed primary human foreskin fibroblasts (HFFs) (Bakkenist, C.J., et al. (2004). Cancer Res. 64(11):3748-3752).
Western Blotting Analysis: A representative lot detected epirubicin-induced ATM Ser1981 (Ser1987 in mouse) phosphorylation in mouse embryonic fibroblasts (MEFs) and human MCF-7 cells (Khongkow, P., et al. (2014). Oncogene. 33(32): 4144-4155).
Western Blotting Analysis: A representative lot detected camptothethin-induced ATM Ser1987 (Ser1981 in human) phosphorylation in primary mouse cortical neurons (Brochier, C., et al. (2013). J. Neurosci. 33(20): 8621-8632).
Western Blotting Analysis: A representative lot detected radiation-/IR-induced ATM Ser1981 phosphorylation in HT29, HeLa, and HEK293T cells (Bardelle, C., and Boros, J. (2012). J. Biomol. Screen. 17(7):912-920).
Western Blotting Analysis: A representative lot detected ionizing radiation-/IR-induced ATM Ser1981 phosphorylation in U2OS cells by Western blotting using whole cell lyates or B56γ immunoprecipitate (Shouse, G.P., et al. (2011). Oncogene. 30(35):3755-3765).
Western Blotting Analysis: A representative lot detected ionizing radiation-/IR-induced ATM Ser1981 phosphorylation in cultured primary human foreskin fibroblasts (HFFs), as well as passage-dependent basal ATM Ser1981 phosphorylation levels in HFFs (Bakkenist, C.J., et al. (2004). Cancer Res. 64(11):3748-3752).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Cell Cycle, DNA Replication & Repair

Reacts with ATM Kinase phosphorylated at serine 1981. Clone 10H11.E12 is covered by US patent No. 6,916,627 and 7,108,992.

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Evaluated by Immunocytochemistry in HeLa cells.

Immunocytochemistry Analysis: 4.0 µg/mL of this antibody detected Camptothecin (Cat. No. 208925) treatment-induced increase of nuclear ATM pSer1981 foci in HeLa cells.

Concentration: Please refer to lot specific datasheet.

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