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9555
cdc25C Antibody Sampler Kit

cdc25C Antibody Sampler Kit #9555

 Image 1

Western blot analysis of extracts from HeLa, 293 and K-562 cells using cdc25C (5H9) Rabbit mAb #4688.

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 Image 2

Western blot analysis of extracts from HT-29 cells, untreated, nocodazole-treated or λ phosphatase-treated, using Phospho-cdc25C (Ser216) Rabbit mAb #4901 (upper) and cdc25C (5H9) Rabbit mAb #4688 (lower).

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 Image 3

Western blot analysis of extracts from HT-29 cells, untreated, nocodazole-treated or λ phosphatase-treated, using Phospho-cdc25C (Thr48) #9527 (upper) and cdc25C (5H9) Rabbit mAb #4688 (lower).

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Western Blotting Image 4

Western Blot analysis of HT29 cell extracts untreated, nocodazole-treated and lambda phosphotase-treated using Phospho-cdc25C (Thr48) (upper), and cdc25C (5H9) Rabbit mAb, #4688 (lower).

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Western Blotting Image 5

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).

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Western Blotting Image 6

Western blot analysis of extracts from HeLa, 293 and K562 cells, using cdc25C (5H9) Rabbit mAb.

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Western Blotting Image 7

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 8

Western blot analysis of GST-cdc25C, unphosphorylated or phosphorylated in vitro by Chk2, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb (upper) or cdc25C antibody #9522 (lower).

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Western Blotting Image 9

Western blot analysis of HT29 cell extracts untreated, nocodazole-treated, and λ phosphotase-treated using Phospho-cdc25C (Thr48), #9527 (upper), and cdc25C (5H9) Rabbit mAb (lower).

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb.

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic localization, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human lymph node, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb in the presence of control peptide (left) or Phospho-cdc25C (Ser216) Blocking Peptide #1190 (right).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-cdc25C (Thr48) Antibody 9527 20 µl
  • WB
H 75 Rabbit 
Phospho-cdc25C (Ser216) (63F9) Rabbit mAb 4901 20 µl
  • WB
  • IP
  • IHC
H Mk 60 Rabbit IgG
cdc25C (5H9) Rabbit mAb 4688 20 µl
  • WB
H 60, 75 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The cdc25C Antibody Sampler Kit provides an economical means to investigate the entry of eukaryotic cells into mitosis. The kit contains enough primary and secondary antibodies to perform two Western blots with each antibody.

Each antibody in the cdc25C Antibody Sampler Kit detects endogenous levels of its respective target protein and does not cross-react with other family members.

Monoclonal antibody is produced by immunizing animals with full-length cdc25C protein or by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser216 of human cdc25C. 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.

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).

  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. Izumi, T. and Maller, J.L. (1993) Mol. Biol. Cell 4, 1337-1350.
  7. Stukenberg, P. T. et al. (2001) Mol. Cell 7, 1071-1083.
  8. Yaffe, M. B. et al. (1997) Science 278, 1957-1960.
  9. Lu, P. J. et al. (1999) Science 283, 1325-1328.
  10. Landrieu, I. et al. (2001) J. Biol. Chem 276, 1434-1438.
Entrez-Gene Id
995
Swiss-Prot Acc.
P30307
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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