Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

cdc25A Antibody #3652

Applications Reactivity Sensitivity MW (kDa) Source
W H M Endogenous 70 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

cdc25A Antibody recognizes endogenous levels of total cdc25A protein. The antibody will cross-react with calf intestinal phosphatase (CIP) when present in abundance.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser177 of human cdc25A.

Western Blotting

Western Blotting

Western blot analysis of extracts from asynchronous (lane 1) or synchronized (lanes 2-4) NIH/3T3 cells, using Phospho-cdc25A (Thr506) Antibody #3654 (upper) or cdc25A Antibody (lower). Cells were synchronized by mitotic shake-off and replated for the indicated times.

Background

The cdc25 protein phosphatase family plays a critical role in activating cyclin-dependent kinases (CDKs) via dephosphorylation of conserved Thr14/Tyr15 inhibitory phosphorylation sites. While cdc25C is primarily responsible for activating CDK1 to overcome the G2/M checkpoint and allow mitotic entry, the primary substrate of cdc25A is CDK2, which, when active, allows progression through the G1/S and intra-S checkpoints (1). Abundance, subcellular localization and activity of cdc25A is tightly controlled by a variety of mechanisms, including phosphorylation, ubiquitination, and inhibitory binding to 14-3-3 proteins. During normal cell cycle progression, elevated c-Myc and E2F transcription factor levels lead to increased cdc25A expression (2). When conditions are favorable for DNA synthesis, cdc25A and CDK2 form an activation loop, wherein each activates the other enzyme (1). DNA damage, on the other hand, leads to multisite phosphorylation at inhibitory sites (Ser123, Ser177, Ser278, Ser292, and Thr506) by Chk1 and Chk2, which result in 14-3-3 binding and ubiquitin-mediated degradation (3,4).

  1. Hoffmann, I. et al. (1994) EMBO J. 13, 4302-4310.
  2. Vigo, E. et al. (1999) Mol. Cell. Biol. 19, 6379-6395.
  3. Chen, M. et al. (2003) Mol. Cell. Biol. 23, 7488-7497.
  4. Sorensen, C.S. et al. (2003) Cancer Cell 3, 247-258.

Application References

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Companion Products

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

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