Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-cdc25C (Ser198) Antibody #9529

Applications Reactivity Sensitivity MW (kDa) Source
W F H Transfected Only 75 Rabbit

Applications Key:  W=Western Blotting  F=Flow Cytometry
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 (Ser198) Antibody detects levels of cdc25C only when phosphorylated at Ser198 and only when derived from a transfected DNA construct.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser198 of human cdc25C. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of GST-cdc25C, unphosphorylated or phosphorylated by HTScan® PLK1 Kinase #7611, using Phospho-cdc25C (Ser198) Antibody (upper) or GST (26H1) Mouse mAb #2624 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells overexpressing full-length GST-cdc25C, untreated or treated with calyculin A #9902, using Phospho-cdc25C (Ser198) Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Phospho-cdc25C (Ser198) Antibody versus propidium iodide (DNA content). The boxed population indicates phospho-cdc25C (Ser198)-positive cells.


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).

During prophase, polo-like kinase 1 (PLK1) phosphorylates cdc25C at Ser198, causing translocation from the cytoplasm to the nucleus, where cdc25C can interact with cdc2/cyclin B to allow for progression through the remaining stages of mitosis (6).

  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.
  6. Toyoshima-Morimoto, F. et al. (2002) EMBO Rep. 3, 341-348.

Application References

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Companion Products

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

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