Phospho-(Ser/Thr) ATM/ATR Subs

2851:

ELISA Peptide , IHC / Paraffin , Immunoprecipitation , Western Blotting

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.)

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

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 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

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)

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

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.)

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.

• Schwartz, M.F. et al. (2002) Mol Cell 9, 1055-65. Applications: Western Blotting
• DiTullio, R.A. et al. (2002) Nat Cell Biol 4, 998-1002. Applications: IC-IF
• Ismail, I.H. et al. (2004) Oncogene 23, 873-82. Applications: Western Blotting
• Lou, Z. et al. (2003) J Biol Chem 278, 13599-602. Applications: IC-IF
• Lindström, M.S. and Wiman, K.G. (2003) Oncogene 22, 4993-5005. Applications: IC-IF
• Brumbaugh, K.M. et al. (2004) Mol Cell 14, 585-98. Applications: IP Western Blotting
• d'Adda di Fagagna, F. et al. (2003) Nature 426, 194-8. Applications: IC-IF Western Blotting
• Demonacos, C. et al. (2004) Nat Cell Biol 6, 968-76. Applications: Western Blotting
• Uziel, T. et al. (2003) EMBO J 22, 5612-21. Applications: IC-IF
• Lee, S.J. et al. (2003) Mol Cell Biol 23, 6300-14. Applications: Western Blotting
• Izumi, N. et al. (2012) Cancer Sci 103, 50-7. Applications: Western Blotting

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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.