Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Chk2 (1C12) Mouse mAb #3440

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IF-IC H Mk Endogenous 62 Mouse IgG2b

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
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

Specificity / Sensitivity

Chk2 (1C12) Mouse mAb detects endogenous levels of total Chk2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with truncated recombinant GST-Chk2.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Chk2 (1C12) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Chk2 (1C12) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Chk2 (1C12) Mouse mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Chk2 (1C12) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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).

  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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For Research Use Only. Not For Use In Diagnostic Procedures.

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