Cell Signaling Technology

Product Pathways - DNA Damage

Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb #2197

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P F H M Hm (Mk) Endogenous 62 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  Mk=Monkey  Hm=Hamster
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb detects endogenous levels of Chk2 only when phosphorylated at Thr68.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Thr68 of human Chk2.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or UV-treated, using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IP

IP

Immunoprecipitation of phospho-chk2 from UV-treated HT29 cells using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb followed by western blot using the same antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 cell pellets, control (left) or UV-treated (right), using Phospho-Chk2 (Thr68) (C13C1) Rabbit mAb.


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

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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