Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Cyclin B1 (V152) Mouse mAb (Alexa Fluor® 488 Conjugate) #4112

Applications Reactivity Sensitivity Isotype
F H M (Hm) Endogenous Mouse IgG1

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

Protocols

Specificity / Sensitivity

Cyclin B1 (V152) Mouse mAb detects endogenous levels of cyclin B1 independent of phosphorylation.

Source / Purification

Cyclin B1 (V152) Monoclonal antibody is produced by immunizing animals with a peptide corresponding to a sequence from hamster cyclin B1. The antibody was conjugated to Alexa Fluor®488 under optimal conditions with an F/P ratio of 2-5.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Cyclin B1 (V152) (Alexa Fluor®488 Conjugate) Mouse mAb versus propidium iodide (DNA content).

Description

Cell Signaling Technology Antibody conjugated to Alexa Fluor ® 488 fluorescent dye and tested in-house for direct Flow Cytometric analysis of human cells. The unconjugated antibody, #4135, reacts with human and mouse. CST expects that Cyclin B1 (V152) Mouse mAb (Alexa Fluor®488 conjugate) will also recognize Cyclin B1 in these species.

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.
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  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.

Application References

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Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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