Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Cell Cycle / Checkpoint

MCM2 (D7G11) XP® Rabbit mAb #3619

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC ChIP H M R Mk Endogenous 125 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  ChIP=Chromatin IP
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

Specificity / Sensitivity

MCM2 (D7G11) XP® Rabbit mAb detects endogenous levels of total MCM2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues of human MCM2.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using MCM2 (D7G11) XP® Rabbit mAb.

IP

IP

Immunoprecipitation of MCM2 from HeLa cell lysates using MCM2 (D7G11) XP® Rabbit mAb followed by western blot using the same antibody. Lane 1 is 5% input.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using MCM2 (D7G11) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using MCM2 (D7G11) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human small intestine using MCM2 (D7G11) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using MCM2 (D7G11) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes, and either 20 μl of MCM2 (D7G11) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human TAP1 Promoter Primers #5148, human IRF-1 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

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). 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 3,9).

  1. Lei, M. and Tye, B.K. (2001) J Cell Sci 114, 1447-54.
  2. Lygerou, Z. and Nurse, P. (2000) Science 290, 2271-3.
  3. Forsburg, S.L. (2004) Microbiol Mol Biol Rev 68, 109-31.
  4. Tye, B.K. and Sawyer, S. (2000) J Biol Chem 275, 34833-6.
  5. Labib, K. et al. (2000) Science 288, 1643-7.
  6. Charych, D.H. et al. (2008) J Cell Biochem 104, 1075-86.
  7. Masai, H. et al. (2006) J Biol Chem 281, 39249-61.
  8. Lin, D.I. et al. (2008) Proc Natl Acad Sci USA 105, 8079-84.
  9. Bailis, J.M. et al. (2008) Mol Cell Biol 28, 1724-38.

Application References

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

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