Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb #4910

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H (M) (R) (Mk) (X) (Z) (B) Endogenous 95 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  Z=Zebrafish  B=Bovine
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-Wee1 (Ser642) (D47G5) Rabbit mAb detects endogenous levels of wee1 protein only when phosphorylated at Ser642.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser642 of human wee1.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 and H441 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb (upper) or Wee1 Antibody #4936 (lower).

IP

IP

Immunoprecipitation of phospho-wee1 from A431 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb followed by Western blot using the same antibody.

Background

Entry of all eukaryotic cells into mitosis is regulated by activation of cdc2 kinase. The critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of Tyr15 and Thr14 (1,2). Phosphorylation at Tyr15 and Thr14 and inhibition of cdc2 is carried out by Wee1 and Myt1 protein kinases, while Tyr15 dephosphorylation and activation of cdc2 is carried out by the cdc25 phosphatase (1,3,4). Hyperphosphorylation and inactivation of Myt1 in mitosis suggests that one or more kinases activated at the G2/M transition negatively regulates Myt1 activity. Kinases shown to phosphorylate Myt1 include cdc2, p90RSK, Akt, and Plk1 (5-8).

Akt/PKB-dependent phosphorylation at Ser642 promotes a change in Wee1 localization from nuclear to cytoplasmic and is associated with G2/M arrest (9).

  1. Watanabe, N. et al. (1995) EMBO J. 14, 1878-1891.
  2. Hunter, T. (1995) Cell 80, 225-236.
  3. Galaktionov, K. et al. (1995) Genes Dev. 9, 1046-1058.
  4. McGowan, C.H. and Russell, P. (1993) EMBO J. 12, 75-85.
  5. Booher, R. N. et al. (1997) J. Biol. Chem. 272, 22300-22306.
  6. Palmer, A. et al. (1998) EMBO J. 17, 5037-5047.
  7. Okumura, E. et al. (2002) Nat. Cell Biol. 4, 111-116.
  8. Nakajima, H. et al. (2003) J. Biol. Chem. 278, 25277-25280.
  9. Katayama, K. et al. (2005) Mol Cell Biol 25, 5725-37.

Application References

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

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