Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

MRN Complex Antibody Sampler Kit #8344

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Mre11 (31H4) Rabbit mAb #4847 40 µl W IP IHC-P IHC-F F H 81 Rabbit IgG
Phospho-Mre11 (Ser676) Antibody #4859 40 µl W H (Mk) 81 Rabbit
Rad50 Antibody #3427 40 µl W H Mk 153 Rabbit
p95/NBS1 Antibody #3002 40 µl W H M R Mk Mi 95 Rabbit
Phospho-p95/NBS1 (Ser343) Antibody #3001 40 µl W E-P H M Mi 95 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  F=Flow Cytometry  E-P=ELISA (Peptide)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Mi=Mink
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Antibodies detect endogenous levels of their respective proteins.

Western Blotting

Western Blotting

Western blot analysis of extracts from Mv 1 Lu cells, treated with UV or hydroxyurea (HU) for the indicated times, using Phospho-p95/NBS1 (Ser343) Antibody #3001.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using p95/NBS1 Antibody #3002.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and K562 cells using Rad50 Antibody #3427.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and K-562 cells using Mre11 (31H4) Rabbit mAb #4847.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and HT-1080 cells, untreated or treated with UV, using Phospho-Mre11 (Ser676) Antibody #4859 (upper) or total Mre11 Antibody #4895 (lower).

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 four western blot experiments with each primary antibody.

Source / Purification

Total polyclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of human Rad50 or carboxy-terminal residues of human p95/NBS1. Activation state-specific polyclonal antibodies are produced by immunizing rabbits with synthetic phosphopeptides corresponding to residues surrounding Ser343 of human p95/NBS1 or Ser676 of human Mre11. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys496 of human Mre11A.

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.

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