Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

CDT1 (D10F11) Rabbit mAb #8064

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Mk Endogenous 65 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  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

CDT1 (D10F11) Rabbit mAb recognizes endogenous levels of total CDT1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human CDT1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CDT1 (D10F11) Rabbit mAb.

IP

IP

Immunoprecipitation of CDT1 from HeLa cells using CDT1 (D10F11) Rabbit mAb. Western blot was performed using the same antibody. Lane 1 is 10% input.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-29 and HUVEC cells using CDT1 (D10F11) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

The initiation of DNA replication in mammalian cells is a highly coordinated process that ensures duplication of the genome only once per cell division cycle. Origins of replication are dispersed throughout the genome, and their activities are regulated via the sequential binding of prereplication and replication factors. The origin recognition complex (ORC) is thought to be bound to chromatin throughout the cell cycle (1,2). The prereplication complex (Pre-RC) forms in late mitosis/early G1 phase beginning with the binding of CDT1 and cdc6 to the origin, which allows binding of the heterohexameric MCM2-7 complex. The MCM complex is thought to be the replicative helicase, and formation of the pre-RC is referred to as chromatin licensing. Subsequent initiation of DNA replication requires the activation of the S-phase promoting kinases CDK2 and cdc7. Cdc7, which is active only in complex with its regulatory subunit dbf4, phosphorylates MCM proteins bound to chromatin and allows binding of the replication factor cdc45 and DNA polymerase (3,4).Binding of CDT1 to geminin prevents pre-RC formation, and expression and degradation of geminin serve to regulate CDT1 activity (reviewed in 5). The interaction of CDT1 with MCM proteins is important in pre-RC formation and licensing (6,7). Both cdc6 and CDT1 are degraded by the ubiquitin proteasome pathway in response to DNA damage associated with rereplication (8).

  1. Okuno, Y. et al. (2001) EMBO J 20, 4263-77.
  2. McNairn, A.J. et al. (2005) Exp Cell Res 308, 345-56.
  3. Bell, S.P. and Dutta, A. (2002) Annu Rev Biochem 71, 333-74.
  4. Tsuji, T. et al. (2006) Mol Biol Cell 17, 4459-72.
  5. Tada, S. (2007) Front Biosci 12, 1629-41.
  6. You, Z. and Masai, H. (2008) J Biol Chem 283, 24469-77.
  7. Teer, J.K. and Dutta, A. (2008) J Biol Chem 283, 6817-25.
  8. Hall, J.R. et al. (2008) J Biol Chem 283, 25356-63.

Application References

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

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