Mre11 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13219
Key Product Details
Species
Applications
Product Summary for Mre11 Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC209414
Accession#: NM_005591
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Product Specifications
Application Notes
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
TMW
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Type
Scientific Data Images for Mre11 Overexpression Lysate
Western Blot: Mre11 Overexpression Lysate [NBL1-13219]
Western Blot: Mre11 Overexpression Lysate [NBL1-13219] - (Left image) - Empty vector transfected control cell lysate (HEK293 cell lysate); (Right image) -Over-expression Lysate for Mre11. A band appears at the theoretical molecular weight of 80.4 kDa for the Over-expression Lysate analysis.Formulation, Preparation, and Storage
Formulation
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Background: Mre11
The MRN assembles as a hetero-hexamer consisting of 2 RAD50 subunits, 2 MRE11 nucleases, and 2 NBS1 subunits, specific to eukaryotes. Mre11 is highly conserved with the N-terminal region of the human protein sharing approximately 50% amino acid sequence identity with the yeast ortholog. Human Mre11 has 3 known isoforms with the predominant isoform having a theoretical molecular weight of 80.6kDa. This nuclear localized protein has high expression in proliferating tissues. Defects in Mre11 can lead to genomic instability, telomere shortening, aberrant meiosis, hypersensitivity to DNA damage, and ataxia telangiectasia-like disorder (ATLD). MRN expression has been associated with the prognosis of cancer and MRN mutants have been linked to cancer susceptibility in ovarian cancer and glioma (4, 5).
References
1. Paull TT. (2018) 20 Years of Mre11 Biology: No End in Sight. Mol Cell. 71(3):419-427. PMID: 30057197
2. Syed A, Tainer JA. (2018) The MRE11-RAD50-NBS1 Complex Conducts the Orchestration of Damage Signaling and Outcomes to Stress in DNA Replication and Repair. Annu Rev Biochem. 87:263-294. PMID: 29709199
3. Wang Y, Cortez D, Yazdi P, Neff N, Elledge SJ, Qin J. (2000) BASC, a super complex of BRCA1-associated proteins involved in the recognition and repair of aberrant DNA structures. Genes Dev. 14(8):927-39. PMID: 10783165
4. Kim JH, Grosbart M, Anand R, Wyman C, Cejka P, Petrini JHJ. (2017) The Mre11-Nbs1 Interface Is Essential for Viability and Tumor Suppression. Cell Rep. 18(2):496-507. PMID: 28076792
5. Bian L, Meng Y, Zhang M, Li D. (2019) MRE11-RAD50-NBS1 complex alterations and DNA damage response: implications for cancer treatment. Mol Cancer. 18(1):169. PMID: 31767017
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Additional Mre11 Products
Product Documents for Mre11 Overexpression Lysate
Product Specific Notices for Mre11 Overexpression Lysate
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.