Cell Signaling Technology

Product Pathways - DNA Damage

Phospho-Chk1(Ser280) Antibody #2347

Applications Reactivity MW (kDa) Source
W H 56 Rabbit

Applications Key:  W=Western Blotting
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-Chk1 (Ser280) Antibody detects endogenous levels of Chk1 only when phosphorylated at serine 280.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser280 of human Chk1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, untreated or UV-treated, using Phospho-Chk1 (Ser280) Antibody (upper) or Chk1 Antibody #2345 (lower).

Background

Chk1 kinase acts downstream of ATM/ATR kinase to play an important role in DNA damage checkpoint control, embryonic development and tumor suppression (1). Activation of Chk1 involves phosphorylation of Ser317 and Ser345 and occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). Chk1 is also phosphorylated at Ser280 and Ser296 following DNA damage. Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (3). Chk1 can also phosphorylate p53 at Ser20 in vitro (4).

Akt has been shown to phosphorylate Chk1 at Ser280 in vitro, and preliminary data suggests that Akt activity may prevent Chk1 activation (5).

  1. Martinho, R.G. et al. (1998) EMBO J. 17, 7239-7249.
  2. Zhao, H. et al. (2001) Mol. Cell. Biol. 21, 4129-4139.
  3. Zeng, Y. et al. (1998) Nature 395, 507-510.
  4. Shieh, S. et al. (2000) Genes Dev. 14, 289-300.
  5. Shtivelman, E. et al. (2002) Curr. Biol. 12, 919-924.

Application References

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