Cell Signaling Technology

Product Pathways - Screening Technologies

Phospho-ATM/ATR Substrate (S*Q) (D23H2/D69H5) Rabbit mAb #9607

Applications Reactivity Sensitivity Isotype
W IP All Endogenous Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-ATM/ATR Substrate Motif (S*Q) (D23H2/D69H5) Rabbit mAb recognizes peptides and proteins containing sequences of phospho-Ser followed by Gln at the +1 position. The antibody does not cross-react with corresponding nonphosphorylated sequences or with other phospho-Ser containing motifs.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing the S*Q motif sequence. The antibody is formulated from two rabbit monoclones in order to cover a broad range of reactivity.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or UV-treated (+, 2 hr), using Phospho-ATM/ATR Substrate (S*Q) (D23H2/D69H5) Rabbit mAb. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biotechnologies).

IP

IP

Immunoprecipitation of HeLa cells, untreated (-) or UV-treated (+, 2 hr) (lanes 3 and 4), using Phospho-ATM/ATR Substrate (S*Q) (D23H2/D69H5) Rabbit mAb. 10% input is shown in lanes 1 and 2. Western blot analysis was performed using the same antibody (upper) and Chk1 (2G1D5) Mouse mAb #2360 (lower).

Background

Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are related kinases that regulate cell cycle checkpoints and DNA repair (1). The identified substrates for ATM are p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1, and Chk1 (1,2) The essential requirement for the substrates of ATM/ATR is S*/T*Q. 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*/T*Q are negative determinants for substrate phosphorylation (3). The complex phenotype of AT cells suggests that it likely has additional substrates (3). To better understand the kinase and identify substrates for ATM and the related kinase ATR, CST has developed antibodies that recognize phosphorylated serine or threonine in the S*/T*Q motif.

  1. Kastan, M.B. and Lim, D.S. (2000) Nature Rev. Mol. Cell Biol. 1, 179-186.
  2. Zhao, H. and Piwnica-Worms, H. (2001) Mol. Cell. Biol. 21, 4129-4139.
  3. Kim, S. T. et al. (1999) J. Biol. Chem. 274, 37538-37543.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.’s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

For Research Use Only. Not For Use In Diagnostic Procedures.

Products