Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Wee1 Antibody #4936

Applications Reactivity Sensitivity MW (kDa) Source
W IP F H R Mk Endogenous 95 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  F=Flow Cytometry
Reactivity Key:  H=Human  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Wee1 Antibody detects endogenous levels of Wee1 protein independent of phosphorylation.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino-terminus of human Wee1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or Lambda Phosphatase-treated NEB #P0753 (10,000 units/ml for 1h), using Wee1 Antibody.

IP

IP

Western blot analysis, using Wee1 Antibody, of extracts from HeLa and 293 cells (lanes 1 and 3) and of protein immunoprecipitated with Wee1 Antibody from the same lysates (lanes 2 and 4).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Wee1 Antibody versus propidium iodide (DNA content). The boxed population indicates Wee1-positive cells.


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).

Wee1 is inactivated upon mitotic entry by phosphorylation at Ser53 and Ser123 by Plk1 and cdc2, followed by beta-TrCP-mediated ubiquitination and degradation (1,9,10).

  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. Parker, L. L. et al. (1995) Proc. Natl. Acad. Sci. U S A 92, 9638-9642.
  10. Watanabe, N. et al. (2004) Proc. Natl. Acad. Sci. USA 101, 4419-4424.

Application References

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Companion Products

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

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