Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Cyclin B1 (Ser147) Antibody #4131

Applications Reactivity Sensitivity MW (kDa) Source
W H M R (Pg) Endogenous 60 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Pg=Pig
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Cyclin B1 (Ser147) Antibody detects endogenous levels of cyclin B1 only when phosphorylated at serine 147.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser147 of human cyclin B1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, using Phospho-Cyclin B1 (Ser147) Antibody (A) and Cyclin B1 (V152) Monoclonal Antibody (C). Treatment of the membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer abolishes the phospho-cyclin B1 signal (B), but has no effect on the total cyclin B1 signal (D).

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, untreated (cont.) or treated with 50 ng/ml nocodazole (M), using Phospho-cyclin B1 (Ser147) Antibody (upper), Cyclin B1 (V152) Monoclonal Antibody (middle), or cdc25C Antibody #9522 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from asynchronous cells (lane 1), S-phase and G2/M blocked cells (lane 2), or synchronized cells collected at various time points following release from this block (lanes 3-7) using Phospho-Cyclin B1 (Ser147) Antibody (upper). Cell cycle synchronization was verified by flow cytometric analysis of DNA content (lower). (Data kindly provided by Ethan Kohn and Alan Eastman, Dartmouth Medical School, Hanover, NH).


Background

Entry of all eukaryotic cells into mitosis is regulated by activation of cdc2 kinase. Activation of cdc2 is controlled at several steps including cyclin B1 binding, phosphorylation of cdc2 at Thr161 and dephosphorylation of cdc2 at Thr14/Tyr15 (1-5). The protein levels of CDK inhibitors and the CDK-associated cyclins are regulated by phosphorylation, ubiquitination and degradation, allowing for a stoichiometric regulation of cell cycle events (6). Four cyclin B1 phosphorylation sites (Ser126, 128, 133 and 147) are located in the cytoplasmic retention signal (CRS) domain and are thought to regulate the translocation of cyclin B1 to the nucleus at the G2/M checkpoint (8-10). Phosphorylation of cyclin B1 is required for cdc25C-dependent dephosphorylation of Tyr15 within cdc2 and subsequent cdc2/cyclin B1 activation (11). While cdc2/cyclin B1 itself can phosphorylate Ser126 and Ser128 (8), polo-like kinase 1 phosphorylates cyclin B1 preferentially at Ser133 and possibly at Ser147 as well (11-13).

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  2. Atherton-Fessler, S. et al. (1993) Mol. Cell. Biol. 13, 1675-1685.
  3. Watanabe, N. et al. (1995) EMBO J. 14, 1878-1891.
  4. Galaktionov, K. et al. (1995) Genes Dev. 9, 1046-1058.
  5. Hunter, T. et al. (1995) Cell 80, 225-236.
  6. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.
  7. McGowan, C.H. et al. (1993) EMBO J. 12, 75-85.
  8. Izumi, T. et al. (1991) Mol. Cell. Biol. 11, 3860-3867.
  9. Li, J. et al. (1995) Mol. Biol. Cell 6, 1111-1124.
  10. Li, J. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 502-507.
  11. Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-220.
  12. Peter, M. et al. (2002) EMBO Rep. 3, 551-556.
  13. Jackman, M. et al. (2003) Nat. Cell Biol. 5, 143-148.

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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