Analysis Note

ControlIrradiated HeLa cell lysates

Application

Use Anti-phospho-ATM (Ser1981) Antibody, clone 10H11.E12 (Mouse Monoclonal Antibody) validated in ICC, IF, IP, WB to detect phospho-ATM (Ser1981) also known as A-T mutated.

Immunoprecipitation:
Phosphorylated ATM was immunoprecipitated from irradiated HeLa cells (Figure A, lanes 3 and 4).

Immunocytochemistry:
Foci are detected in irradiated human and mouse fibroblasts. Determined by an independent laboratory.

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryCell Cycle, DNA Replication & Repair

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

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). The identified substrates for ATM are p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1 and Chk1. The essential requirement for the substrates of ATM/ATR is S/TQ. Hydrophobic amino acids at positions -3 and -1, and negatively charged amino acids at position +1 are positive determinants for substrate recognition by these kinases. Positively charged residues surrounding the S/TQ are negative determinants for substrate phosphorylation. The complex phenotype of cells derived from patients with AT suggests that ATM has additional cellular substrates. In unirradiated cells, ATM is present as an inactive homodimer or multimer. Double-stranded breaks in DNA caused by ionizing radiation cause rapid ATM kinase activation through dissociation of this complex and ATM autophosphorylation at Ser1981.

Immunogen

KLH-conjugated, synthetic peptide corresponding to amino acids 1974-1988 (SLAFEEG[pS]QSTTISS) of human ATM. The immunizing sequence has 11/12 identical amino acids in mouse and rat.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Format: Purified

Protein G purified mouse IgG in 0.014 M phosphate buffer, pH 7.6, with 0.175 M NaCl, 0.07 % Sodium Azide and 30% glycerol. Liquid at -20°C.

Quality

Routinely evaluated by immunoblot on in crude lysates from irradiated HeLa cells.

Western Blot Analysis:
0.5 µg/mL of this lot detected phosphorylated ATM in crude lysates from irradiated HeLa cells.

Specificity

This antibody recognizes ATM, Mr ~370 kDa. A non-specific protein was also detected, Mr ~ >400 kDa.

Predicted to cross-react with rat based on sequence homology

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

~370 kDa