Product Pathways - DNA Damage
Phospho-Chk2 (Ser33/35) Antibody #2665
PhosphoSitePlus® protein, site, and accession data: Chk2
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | H Mk | Endogenous | 62 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 2665:
- Western Blotting
Specificity / Sensitivity
Phospho-Chk2 (Ser33/35) Antibody detects endogenous levels of Chk2 only when phosphorylated at serine 33/35. The antibody does not cross-react with Chk2 phosphorylated at other sites.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser33/35 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 (Ser33/35) Antibody.
Western Blotting
Western blot analysis of extracts from HeLa cells treated with UV for the indicated times, using Phospho-Chk2 (Ser33/35) 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 at 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
- Mochan, T. A. et al. (2003) 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activiating Ataxia-Telangiectasia Mutated (ATM) in response to DNA damage. Cancer Research 63, 8586-8591. Applications: Western Blotting
- Buscemi, G. et al. (2006) Mol Cell Biol 26, 7832-45. Applications: Western Blotting
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
- 2662 Chk2 Antibody
- 2661 Phospho-Chk2 (Thr68) Antibody
- 2666 Phospho-Chk2 (Ser19) Antibody
- 2669 Phospho-Chk2 (Ser516) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
