Q: Could you recommend an optimal Mre11 positive control for Western blot among your products?

A: NBL1-13219 would be the best product since it is intended for this purpose.

Mre11 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-13219

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Mre11 Overexpression Lysate

Mre11 Transient 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

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

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.

Theoretical MW

80.4 kDa.
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

Overexpression

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

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Mre11

Mre11 (double-strand break repair protein MRE11A) is an important component of MRN/X complex (Mre11, Rad50, and Nbs1/Xrs2), playing a central role in double-strand break (DSB) repair, DNA recombination, telomere integrity maintenance, meiotic cell division, and viral infection. Mre11 and Rad50 comprise the catalytic component of the MRN complex with Rad50 involved in ATP hydrolysis and Mre11 functioning as a nuclease having both single-strand endonuclease and double-strand-specific 3'-5' exonuclease activity (1). The MRN complex is also involved in DNA damage signaling via activation of ATM kinase and in telomeres, where it may modulate t-loop formation (2). In addition, Mre11 is a component of the BASC complex (BRCA1, MSH2, MSH6, MLH1, ATM, BLM, RAD50, MRE11A and NBN) and interacts with DCLRE1C/Artemis as well as DCLRE1B/Apollo (3).

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

Long Name

Meiotic Recombination 11 Homolog A

Alternate Names

MRE11A

Gene Symbol

MRE11

Additional Mre11 Products

Product Documents for Mre11 Overexpression Lysate

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Certificate of Analysis

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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.

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