Product Pathways - DNA Damage
Phospho-Chk2 (Ser19) Antibody #2666
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W IHC-P | H | 62 | Rabbit |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:
H=Human
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 (Ser19) Antibody detects endogenous levels of Chk2 only when phosphorylated at serine 19. 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 Ser19 of human Chk2. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from COS cells, untransfected (lane 1) or transfected with Wild-type Chk2 (lane 2), Chk2 (S19A) (lane 3), Chk2 (T26S28A) (lane 4), Chk2 (S33S35A) (lane 5) or Chk2 (T68A) (lane 6), using Phospho-Chk2 (Ser19) Antibody.
Western Blotting
Western blot analysis of extracts from HeLa cells treated with UV for the indicated times, using Phospho-Chk2 (Ser19) Antibody.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-Chk2 (Ser19) Antibody.
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).
- Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415.
- Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665.
- Murakami, H. and Okayama, H. (1995) Nature 374, 817-819.
- Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186.
- Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394.
- Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765.
- Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936.
- Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537-30541.
Application References
- 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 (Ser19) Antibody in the following applications: Western Blotting
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