Revision 1

#12079Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

125

Source/Isotype:

Mouse IgG1

UniProt ID:

#P49736

Entrez-Gene Id:

4171

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:400
Immunofluorescence (Immunocytochemistry) 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

MCM2 (1E7) Mouse mAb recognizes endogenous levels of total MCM2 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human MCM2 protein.

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). 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).

  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. Tsuji, T. et al. (2006) Mol Biol Cell 17, 4459-72.
  10. Bailis, J.M. et al. (2008) Mol Cell Biol 28, 1724-38.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#12079

MCM2 (1E7) Mouse mAb

Western Blotting Image 1: MCM2 (1E7) Mouse mAb Expand Image
Western blot analysis of extracts from various cell lines using MCM2 (1E7) Mouse mAb.
Immunoprecipitation Image 1: MCM2 (1E7) Mouse mAb Expand Image
Immunoprecipitation of MCM2 from Jurkat cell extracts using Mouse (G3A1) mAb IgG1 Isotype Control #5415 (lane 2) or MCM2 (1E7) Mouse mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using MCM2 (1E7) Mouse mAb.
Immunohistochemistry Image 1: MCM2 (1E7) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using MCM2 (1E7) Mouse mAb.
Immunohistochemistry Image 2: MCM2 (1E7) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lymph node using MCM2 (1E7) Mouse mAb.
Immunohistochemistry Image 3: MCM2 (1E7) Mouse mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using MCM2 (1E7) Mouse mAb.
Immunofluorescence Image 1: MCM2 (1E7) Mouse mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells using MCM2 (1E7) Mouse mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #8584.