Revision 3

#8344Store at -20C

1 Kit

(5 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Mre11 (31H4) Rabbit mAb 4847 20 µl 81 kDa Rabbit IgG
Phospho-Mre11 (Ser676) Antibody 4859 20 µl 81 kDa Rabbit 
Rad50 Antibody 3427 20 µl 153 kDa Rabbit 
Phospho-p95/NBS1 (Ser343) Antibody 3001 20 µl 95 kDa Rabbit 
p95/NBS1 (D6J5I) Rabbit mAb 14956 20 µl 95 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

MRN Complex Antibody Sampler Kit offers an economical way of detecting each target protein. The kit contains enough primary and secondary antibody to perform two western blot experiments with each primary antibody.

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.

Background

The Mre11-Rad50-Nbs1 (MRN) complex is a key mediator of genome maintenance, playing important roles in meiosis, telomere stability at the ends of chromosomes, and the cellular responses to DNA damage (1-5). Homodimers of the Mre11 and Rad50 subunits form a tetramer core that binds directly to DNA and associates with the Nbs1 subunit (6). The complex functions as a sensor of DNA damage and localizes to DNA double-strand breaks. At these DNA lesions, the MRN complex tethers DNA ends and processes free strands via the endonuclease and exonuclease activities of Mre11. In addition to stimulating both homologous recombination and nonhomologous end joining repair DNA pathways, MRN activates DNA damage checkpoint signaling cascades regulating cell cycle progression. In some contexts, MRN is required for ATM activation and downstream phosphorylation of p53, BRCA1, and Chk2 (7). ATM also phosphorylates Mre11, Rad50, and Nbs1 (also known as p95 and Nibrin). Notably, Nbs1 Ser343 and Mre11 Ser676 are phosphorylated by ATM. Phosphorylation modulates function and association with many mediators, some of which include 53BP1, RPA, hSSB1, TRF2, BRCA1, FANCD2, CtP1, Histone H2AX, MDC1, and WRN helicase. Each subunit is essential for mammalian embryonic development, as mice with homozygous-null mutations in Mre11, Nbs1, or Rad50 are lethal. Furthermore, MRN complex function is required in developing lymphocytes for antigen receptor gene recombination initiated by the Rag-1 and Rag-2 recombinases. In humans, Mre11 and Nbs1 mutations cause chromosomal instability and radiosensitivity and are associated with ataxia-telangiectasia-like disorder (ATLD) and Nijmegen breakage syndrome (NBS), respectively (8). Genomic instability and cancer have been shown to develop in cells with genetic mutations within MRN complex genes.

  1. D'Amours, D. and Jackson, S.P. (2002) Nat Rev Mol Cell Biol 3, 317-27.
  2. van den Bosch, M. et al. (2003) EMBO Rep 4, 844-9.
  3. Ajimura, M. et al. (1993) Genetics 133, 51-66.
  4. Deng, Y. et al. (2009) Nature 460, 914-8.
  5. Lamarche, B.J. et al. (2010) FEBS Lett 584, 3682-95.
  6. Williams, R.S. et al. (2009) Cell 139, 87-99.
  7. Uziel, T. et al. (2003) EMBO J 22, 5612-21.
  8. Zhao, S. et al. (2000) Nature 405, 473-7.

Background References

    Trademarks and Patents

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

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    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 3
    #8344

    MRN Complex Antibody Sampler Kit

    MRN Complex Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using p95/NBS1 (D6J5I) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    MRN Complex Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from 293 cells, untreated or UV-treated (100 mJ/cm2, 1 hr), using Phospho-p95/NBS1 (Ser343) Antibody (upper) or p95/NBS1 (E8M3Q) XP® Rabbit mAb #81234 (lower).
    MRN Complex Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from Jurkat and K562 cells, using RAD50 Antibody.
    MRN Complex Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from HeLa and K562 cells, using Mre11 Rabbit (31H4) mAb.
    MRN Complex Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from HeLa and HT-1080 cells, untreated or treated with UV, using Phospho-Mre11 (Ser676) Antibody (upper) or total Mre11 Antibody #4895 (lower).
    MRN Complex Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    MRN Complex Antibody Sampler Kit: Image 7 Expand Image
    Immunoprecipitation of p95/NBS1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or p95/NBS1 (D6J5I) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using p95/NBS1 (D6J5I) Rabbit mAb.
    MRN Complex Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Mre11 (31H4) Rabbit mAb in the presence of control peptide (left) or Mre11 Blocking Peptide #1035 (right).
    MRN Complex Antibody Sampler Kit: Image 9 Expand Image
    Confocal immunofluorescent analysis of SK-MEL-28 (left) and GM07166 (right) cells using p95/NBS1 (D6J5I) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054.
    MRN Complex Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Mre11 (31H4) Rabbit mAb.