Product Pathways - DNA Damage

Phospho-Chk2 (Thr68) Antibody #2661

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Applications	Reactivity	MW (kDa)	Source
W IP IF-IC F	H Mk	62	Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Chk2 (Thr68) Antibody detects endogenous levels of Chk2 only when phosphorylated at threonine 68. The antibody does not cross-react with Chk2 phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr68 of human Chk2. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (1-3). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68) each followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4,5). After DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (5-7). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 on residues Thr383 and Thr387 in the activation loop of the kinase domain (8).

1. Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415.
2. Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665.
3. Murakami, H. and Okayama, H. (1995) Nature 374, 817-819.
4. Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186.
5. Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394.
6. Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765.
7. Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936.
8. Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537-30541.

Application References

• Kohn, E. A. et al. (2002) Abrogation of the S phase DNA damage checkpoint results in S phase progression or premature mitosis depending on the concentration of 7-hydroxystaurosporine and the kinetics of Cdc25C activation. J. Biol. Chem. 277, 26553-26564. This article references the use of Phospho-Chk2 (Thr68) Antibody in the following applications: Western Blotting
• Yin, M. B. et al. (2004) Enhanced 7-ethyl-10-hydroxycamptothecin (SN-38) lethality by methylselenocysteine is associated with Chk2 phosphorylation at threonine-68 and down-regulation of cdc6 expression. Molecular Pharmacology 66, 153-160. This article references the use of Phospho-Chk2 (Thr68) Antibody in the following applications: Western Blotting
• Eastman, A. et al. (2002) A novel indolocarbazole, ICP-1, abrogates DNA damage-induced cell cycle arrest and enhances cytotoxicity: similarities and differences to the cell cycle checkpoint abrogator UCN-01. Mol. Cancer Ther. 1, 1067-1078. This article references the use of Phospho-Chk2 (Thr68) Antibody in the following applications: Western Blotting
• Lukas, C. et al. (2003) Distinct spatiotemporal dynamics of mammalian checkpoint regulators induced by DNA damage. Nat. Cell Biol. 5, 255-260. This article references the use of Phospho-Chk2 (Thr68) Antibody in the following applications: IC-IF Western Blotting
• Castedo, M. et al. (2004) The cell cycle checkpoint kinase Chk2 is a negative regulator of mitotic catastrophe. Oncogene 23, 4353-4361. This article references the use of Phospho-Chk2 (Thr68) Antibody in the following applications: Flow Cytometry IC-IF Western Blotting

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