Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser/Thr) ATM/ATR Substrate Antibody #2851

Applications Reactivity Source
W IP IHC-P E-P All Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Phospho-(Ser/Thr) ATM/ATR Substrate Antibody detects endogenous levels of proteins containing the ATM/ATR substrate motif. This antibody preferentially binds peptides and proteins that contain phospho-Ser/Thr preceded by Leu or similar hydrophobic amino acids at the -1 position and followed by Gln at the +1 position. The antibody does not cross-react with corresponding nonphosphorylated sequences or with other phospho-Ser/Thr-containing motifs. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-(Ser/Thr) ATM/ATR substrate peptides (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or UV-treated, using Phospho-(Ser/Thr) ATM/ATR Substrate Antibody.

Western Blotting

Western Blotting

Western blot analysis of yeast cell extracts, untreated or treated with 4-NQO, a potent activator of ATM/ATR orthologues, using Phospho-(Ser/Thr) ATM/ATR Substrate Antibody.

IP

IP

Western blot analysis of immunoprecipitated protein from Chk2-transfected and UV-treated COS cells, using Chk2 antibody for immunoprecipitation and Phospho-(Ser/Thr) ATM/ATR Substrate Antibody for immunoblotting.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 cells, untreated (left) or treated (right) with UV (upper) or doxorubicin (lower), using Phospho-(Ser/Thr) ATM/ATR Substrate Antibody.

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser/Thr) ATM/ATR Substrate Antibody ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* or T* denote phosphorylated serine or threonine.)

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

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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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.

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