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MRE11 Meiotic Recombination 11 Homolog A (S. Cerevisiae) (MRE11A) antibody

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(138), (69), (61), (6), (4), (3), (3), (2), (2), (1), (1), (1)
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Western Blotting (WB), Chromatin Immunoprecipitation (ChIP), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(119), (56), (42), (26), (24), (22), (13), (6), (5), (1), (1), (1), (1)
Pubmed 1 reference available
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Quantity 50 μL
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Immunogen Full length human Mre11 protein.
Purification Whole antisera
Alternative Name MRE11 (MRE11A Antibody Abstract)
Background The hMre11-hRad50-NBS1 protein plays a central role in the human cellularDNA-Damage response, with recent studies indicating that these proteins help linkDNA-damage detection to DNA-repair and cell cycle-checkpoint functions. This proteincomplex has been implicated in the activation of cell cycle-regulatory pathways, due tothe fact that the NBS1 gene mutates in the chromosomal-instability syndrome, Nijmegenbreakdown syndrome (NBS), which is characterized by increased cancer incidence, cellcycle checkpoint defects, and ionizing radiation sensitivity.HMre11-hRad50 foci form in response to DNA double-strand breaks and rely upon aDNA damage-induced signaling pathway. However, hMre11 migrates to sites ofdamage while hRad51 does not localize at these sites. These findings are consistentwith the distinct role of these proteins in DNA repair. Alternate Names: anti-Ataxia-telangiectasia disorder-like antibody, anti-ATLD antibody, anti-MRE11aantibody, anti-MRE11b antibody.
Gene Symbol: MRE11A
Gene ID 4361, 17535
UniProt P49959
Pathways DNA Damage Repair
Application Notes This Mre11 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections (PMID 21279473) Immunoprecipitation and Western Blot. In WB, a band can be seen at ~81 kDa. For ICC/IF, this antibody has been used with methanol-fixed IMR90 primary human fibroblasts. For IP, the suggested working dilution is 3 ul for immunoprecipitation of 3X10^6 cells. Co-IP application has been reported by Ching et al 2012 (PMID: 22190719). Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 24349281)
Recommended dilutions: Chromatin Immunoprecipitation, Flow Cytometry, Immunocytochemistry/Immunofluorescence 1:200, Immunohistochemistry 1:10-1:500, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin 1:10-1:500, Immunoprecipitation 3 µL, Western Blot 1:5000
Protocol Protocol specific for Mre11 Antibody Protocol specific for Mre11 Antibody Immunoprecipitation Procedure
1. For IP reactions, start with extract (whole cell or nuclear) from around 3 million cells prepared in 0.5-1 mL lysis buffer (100 mM NaCl, 10 mM Tris HCl, 5 mM EDTA, 0.5 % nonidet p40).
. Cells are resuspended in lysis buffer, then incubated with rotation about 15 min at 4 degrees C.
. The lysate is then centrifuged 5 min at 14000g to remove insoluble material.
. To cleared lysate, add 1-3 µL of antiserum and incubate on ice for 30 min.
. Collect immune complexes on Protein A Sepharose by adding 25 µL of a 50 % slurry, and incubate with rotation for 1 hour at 4 degrees C.
. The complexes are pelleted gently (5000g for 5-10 sec.) then washed with 1 mL lysis buffer.
. Repeat the wash 2 more times.
. Analyze the immunoprecipitates by SDS PAGE. This antibody works well for IP reactions from both human and mouse cells. The intact complex is stable and can be immunoprecipitated in many common lysis buffers (up to 0.5 M NaCl). Western Blot Procedure
. Run 50 µg of protein on a 4-20 % Tris-glycine mini-gel at 125V for 90 minutes.
. Equilibrate gel, nitrocellulose membrane, Whatman paper, and blotting pads in transfer buffer for 15 minutes.
. Transfer protein to the membrane at 25V for 90 minutes.
. Allow membrane to air-dry.
. Block membrane with 1XPBS/3 % BSA for 1 hour at room temperature (23-27 degrees C).
. Wash membrane twice, for 5 minutes each, with 1XPBS/0.05 % Tween-20 (PBST).
. Incubate membrane with 1:5000 dilution of NB100-142 (anti-hMre11), diluted in 1XPBS/1 % BSA, for 1 hour at room temperature.
. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
. Incubate membrane with goat anti-rabbit IgG-HRP, diluted in 1XPBS/1 % BSA, for 1 hour at room temperature.
. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
. Detect cross-reacting proteins using Renaissance Chemiluminescence Reagent Plus kit from NEN Life Sciences. NOTE: HeLa whole cell extracts were used as a positive control for this antibody.Immunofluorescence Procedure A 5beta in situ extraction method [10mM Pipes, pH 6.8 / 0.2 % Triton X-100 / 100mM MgCl2 / 100mM sucrose/ 10mM EGTA Beta on ice] followed by 4 % paraformaldehyde fixation of tissues works well for immunofluorescence of anti-hMre11 . Please see reference: Franchitto, A, Pichierri, P, Blooms syndrome protein is required for correct relocalization of RAD50/Mre11/nbs1 complex after replication fork arrest. J. of Cell Biology, DOI: 10 (2002)Immunohistochemistry - FFPE sectionsI. Deparaffinization:A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.B. Treat slides with 100 % Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.II. Quench Endogenous Peroxidase:A. Place slides in peroxidase quenching solution: 15-30 minutes.To Prepare 200 mL of Quenching Solution:Add 3 mL of 30 % Hydrogen Peroxide to 200 mL of Methanol.Use within 4 hours of preparationB. Place slides in distilled water: 2 changes for 2 minutes each.III. Retrieve Epitopes:A. Preheat Citrate
Restrictions For Research Use only
Format Liquid
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
Western Blotting (WB) image for anti-MRE11 Meiotic Recombination 11 Homolog A (S. Cerevisiae) (MRE11A) antibody (ABIN151076) Western Blot analysis of Mre11 on 50 ug of HeLa and MEF lysates, using ABIN151076
Immunohistochemistry (IHC) image for anti-MRE11 Meiotic Recombination 11 Homolog A (S. Cerevisiae) (MRE11A) antibody (ABIN151076) MRE11 staining in the human epidermis detected with ABIN151076.
Product cited in: Wang, Cortez, Yazdi et al.: "BASC, a super complex of BRCA1-associated proteins involved in the recognition and repair of aberrant DNA structures." in: Genes & development, Vol. 14, Issue 8, pp. 927-39, 2000 (PubMed).

Catalog No. ABIN151076

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