Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-cdc25C (Ser216) Antibody #9528

Applications Reactivity Sensitivity MW (kDa) Source
W H Endogenous 60 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-cdc25C (Ser216) Antibody detects endogenous levels of cdc25C only when phosphorylated at Ser216.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser216 of human cdc25C. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of wild type GST-cdc25C and mutant GST-cdc25C Ser216Ala fusion proteins, untreated or phosphorylated by Chk2, using Phospho-cdc25C (Ser216) Antibody.

Western Blotting

Western Blotting

Western blot analysis of GST-cdc25C phosphorylated by cdc2/cyclin B (New England Biolabs #P6020), Chk2 or both kinases, using Phospho-cdc25C (Ser216) Antibody (upper) or Phospho-cdc25C (Ser214) antibody (lower).

Western Blotting

Western Blotting

Western Blot analysis of HT29 cells, untreated, nocodazole-treated, and lambda phosphotase-treated, using Phospho-cdc25C (Ser216) antibody (upper), and cdc25C (5H9) Rabbit mAb, #4688, (lower).


Background

Cdc25 is a protein phosphatase responsible for dephosphorylating and activating cdc2, a crucial step in regulating the entry of all eukaryotic cells into mitosis (1). cdc25C is constitutively phosphorylated at Ser216 throughout interphase by c-TAK1, while phosphorylation at this site is DNA damage-dependent at the G2/M checkpoint (2). When phosphorylated at Ser216, cdc25C binds to members of the 14-3-3 family of proteins, sequestering cdc25C in the cytoplasm and thereby preventing premature mitosis (3). The checkpoint kinases Chk1 and Chk2 phosphorylate cdc25C at Ser216 in response to DNA damage (4,5).

  1. Jessus, C. and Ozon, R. (1995) Prog. Cell Cycle Res. 1, 215-228.
  2. Peng, C.Y. et al. (1997) Science 277, 1501-1505.
  3. Kumagai, A. and Dunphy, W.G. (1999) Genes Dev. 13, 1067-1072.
  4. Blasina, A. et al. (1999) Curr. Biol. 9, 1-10.
  5. Furnari, B. et al. (1999) Mol. Biol. Cell 10, 833-845.

Application References

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

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