Phospho-cdc25C (Thr48) Antibody #9527
Filter:
- WB
Supporting Data
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 75 |
SOURCE | Rabbit |
Application Key:
- WB-Western Blotting
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
Phospho-cdc25C (Thr48) Antibody detects endogenous levels of cdc25C only when phosphorylated at Thr48.
Species Reactivity:
Human
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr48 of human cdc25C. Antibodies are purified by protein A and peptide affinity chromatography.
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).
Full activation of cdc25C involves phosphorylation at more than 12 different sites by cdc2/cyclin B and Polo-like kinase, and the activity of Pin1, a peptidyl-prolyl isomerase (PPI) (6,7). Pin1 contains a WW domain that binds phospho-Ser/Thr-Pro sites and a catalytic PPI region that induces a cis/trans isomerization on phospho-Ser/Thr-Pro bonds (8). Thr48 and Thr67 of cdc25C interact directly with the WW domain of Pin1 when these sites are phosphorylated (9). Thr48 phosphorylation also mediates binding to CKS/p13SUC1 (10).
Full activation of cdc25C involves phosphorylation at more than 12 different sites by cdc2/cyclin B and Polo-like kinase, and the activity of Pin1, a peptidyl-prolyl isomerase (PPI) (6,7). Pin1 contains a WW domain that binds phospho-Ser/Thr-Pro sites and a catalytic PPI region that induces a cis/trans isomerization on phospho-Ser/Thr-Pro bonds (8). Thr48 and Thr67 of cdc25C interact directly with the WW domain of Pin1 when these sites are phosphorylated (9). Thr48 phosphorylation also mediates binding to CKS/p13SUC1 (10).
- Jessus, C. and Ozon, R. (1995) Prog. Cell Cycle Res. 1, 215-228.
- Peng, C.Y. et al. (1997) Science 277, 1501-1505.
- Kumagai, A. and Dunphy, W.G. (1999) Genes Dev. 13, 1067-1072.
- Blasina, A. et al. (1999) Curr. Biol. 9, 1-10.
- Furnari, B. et al. (1999) Mol. Biol. Cell 10, 833-845.
- Izumi, T. and Maller, J.L. (1993) Mol. Biol. Cell 4, 1337-1350.
- Stukenberg, P. T. et al. (2001) Mol. Cell 7, 1071-1083.
- Yaffe, M. B. et al. (1997) Science 278, 1957-1960.
- Lu, P. J. et al. (1999) Science 283, 1325-1328.
- Landrieu, I. et al. (2001) J. Biol. Chem 276, 1434-1438.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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