Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb #4539

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F H M R Mk Endogenous 34 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  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

Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb detects endogenous levels of cdc2 protein only when phosphorylated at tyrosine 15. Based on sequence similarity, the antibody may cross-react with CDK2 and CDK3.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr15 of human cdc2.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or hydroxyurea treated for 20 hours, using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of nonphosphorylated GST-cdc2 fusion protein (lane 1), and extracts from SK-N-MC cells treated with hydroxyurea for 20 hours (lane 2), using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb (upper), or cdc2 Antibody #9112 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb versus propidium iodide (DNA content).


IF-IC

IF-IC

Confocal immunofluorescent analysis of asynchronous HeLa cells labeled with Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb (green) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (red).

Background

The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).

  1. Atherton-Fessler, S. et al. (1994) Mol. Biol. Cell. 5, 989-1001.
  2. Norbury, C. et al. (1991) EMBO. J. 10, 3321-3329.
  3. McGowan, C.H. and Russell, P. (1993) EMBO J. 12, 75-85.
  4. Wells, N.J. et al. (1999) J. Cell. Sci. 112, 3361-3371.
  5. Hunter, T. (1995) Cell 80, 225-236.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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

Products