Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Cdc45 (D7G6) Rabbit mAb #11881

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

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

* Product-specific protocol.

Specificity / Sensitivity

Cdc45 (D7G6) Rabbit mAb recognizes endogenous levels of total cdc45 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human cdc45 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Cdc45 (D7G6) Rabbit mAb.

IP

IP

Immunoprecipitation of cdc45 from Jurkat cell extracts, using Cdc45 (D7G6) Rabbit mAb (lane 2) or Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Cdc45 (D7G6) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293 (left) and WI-38 (right) cells using Cdc45 (D7G6) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).


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 pre-replication and replication factors. The origin recognition complex (ORC) is thought to bind to chromatin throughout the cell cycle (1,2). The pre-replication complex (Pre-RC) forms in late mitosis/early G1 phase 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 phosphorylates MCM proteins bound to chromatin and, in conjunction with CDT1, recruits the replication factor cdc45 (3-5). Cdc45 is required for formation of pre-initiation complexes at the G1/S transition and for activation of replication origins. The level of cdc45 protein expression is associated with the proliferative status of the cell or tissue. Terminally differentiated and senescent cells lack cdc45 and highly proliferative cell lines express high levels of cdc45 (6).

  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. Ballabeni, A. et al. (2009) J Biol Chem 284, 3028-36.
  6. Pollok, S. et al. (2007) FEBS J 274, 3669-84.

Application References

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Companion Products

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

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