Product Pathways - Cell Cycle / Checkpoint
Phospho-MCM3 (Ser112) (D3S4M) Rabbit mAb #12686
|12686S||100 µl (10 western blots)||---||In Stock||---|
|12686||carrier free and custom formulation / quantity||email request|
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Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Species predicted to react based on 100% sequence homology: Mouse, Rat, Hamster, Zebrafish, Dog.
Specificity / Sensitivity
Phospho-MCM3 (Ser112) (D3S4M) Rabbit mAb recognizes endogenous levels of MCM3 protein only when phosphorylated at Ser112. This antibody also recognizes a 200 kDa band of unknown origin in both interphase and mitotic cells.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser112 of human MCM3 protein.
Western blot analysis of HeLa cell extracts, untreated (-) or treated with Nocodazole #2190 (100 ng/ml, 24 hr; +), using Phospho-MCM3 (Ser112) (D3S4M) Rabbit mAb (upper), MCM3 (D47B6) Rabbit mAb #4003 (middle) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Immunoprecipitation of Phospho-MCM3 (Ser112) from HeLa cell extracts, untreated (-) or treated with Nocodazole #2190 (100 ng/ml, 24 hr; +), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 3 and 4) or Phospho-MCM3 (Ser112) (D3S4M) Rabbit mAb (lane 5 and 6). Lanes 1 and 2 are 10% input. Western blot analysis was performed using Phospho-MCM3 (Ser112) (D3S4M) Rabbit mAb.
The minichromosome maintenance (MCM) 2-7 proteins are a family of six related proteins required for initiation and elongation of DNA replication. MCM2-7 bind together to form the heterohexameric MCM complex that is thought to act as a replicative helicase at the DNA replication fork (1-5). This complex is a key component of the pre-replication complex (pre-RC) (reviewed in 1). Cdc6 and CDT1 recruit the MCM complex to the origin recognition complex (ORC) during late mitosis/early G1 phase forming the pre-RC and licensing the DNA for replication (reviewed in 2). Licensing of the chromatin permits the DNA to replicate only once per cell cycle, thereby helping to ensure that genetic alterations and malignant cell growth do not occur (reviewed in 3). Phosphorylation of the MCM2, MCM3, MCM4, and MCM6 subunits appears to regulate MCM complex activity and the initiation of DNA synthesis (6-8). CDK1 phosphorylation of MCM3 at Ser112 during late mitosis/early G1 phase has been shown to initiate complex formation and chromatin loading in vitro (8). Phosphorylation of MCM2 at serine 139 by cdc7/dbf4 coincides with the initiation of DNA replication (9). MCM proteins are removed during DNA replication, causing chromatin to become unlicensed through inhibition of pre-RC reformation. Studies have shown that the MCM complex is involved in checkpoint control by protecting the structure of the replication fork and assisting in restarting replication by recruiting checkpoint proteins after arrest (reviewed in 3,10).
- Lei, M. and Tye, B.K. (2001) J Cell Sci 114, 1447-54.
- Lygerou, Z. and Nurse, P. (2000) Science 290, 2271-3.
- Forsburg, S.L. (2004) Microbiol Mol Biol Rev 68, 109-31.
- Tye, B.K. and Sawyer, S. (2000) J Biol Chem 275, 34833-6.
- Labib, K. et al. (2000) Science 288, 1643-7.
- Charych, D.H. et al. (2008) J Cell Biochem 104, 1075-86.
- Masai, H. et al. (2006) J Biol Chem 281, 39249-61.
- Lin, D.I. et al. (2008) Proc Natl Acad Sci USA 105, 8079-84.
- Tsuji, T. et al. (2006) Mol Biol Cell 17, 4459-72.
- Bailis, J.M. et al. (2008) Mol Cell Biol 28, 1724-38.
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For Research Use Only. Not For Use In Diagnostic Procedures.
XP® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.