Anti-Mre11 antibody [12D7] (ab214)

Product name

Anti-Mre11 antibody [12D7]
See all Mre11 primary antibodies
Description

Mouse monoclonal [12D7] to Mre11
Host species

Mouse
Tested applications

Suitable for: Flow Cyt, ICC/IF, IHC-Fr, WB, IP, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Amino acids 182-582 of Mre11 expressed in E. coli.
Positive control
- T24 cells, recombinant fusion protein.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Concentration information loading...
Purity

Protein G purified
Clonality

Monoclonal
Clone number

12D7
Myeloma

NS1
Isotype

IgG1
Light chain type

kappa
Research areas

Compatible Secondaries
Immunohistochemistry kits
- Mouse on Mouse Polymer IHC Kit (ab127055)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Our Abpromise guarantee covers the use of ab214 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt		Use 1-2µg for 10⁶ cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF		1/200 - 1/500. Wash cells with PBS, then wash with PBS containing 0.5% Triton X-100 and fix for 5 min with PBS containing 3% paraformaldehyde. Block cells for 1 h in PBS containing 15% fetal bovine serum (see Robinson et al).
IHC-Fr		1/200.
WB		Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 79 kDa (predicted molecular weight: 79 kDa). (see Robinson et al).
IP		Use a concentration of 1 - 5 µg/ml. (for normal lymphoblastoid cell lines).
IHC-P		Use at an assay dependent concentration.

Function

Component of the MRN complex, which plays a central role in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity and meiosis. The complex possesses single-strand endonuclease activity and double-strand-specific 3'-5' exonuclease activity, which are provided by MRE11A. RAD50 may be required to bind DNA ends and hold them in close proximity. This could facilitate searches for short or long regions of sequence homology in the recombining DNA templates, and may also stimulate the activity of DNA ligases and/or restrict the nuclease activity of MRE11A to prevent nucleolytic degradation past a given point. The complex may also be required for DNA damage signaling via activation of the ATM kinase. In telomeres the MRN complex may modulate t-loop formation.
Involvement in disease

Defects in MRE11A are a cause of ataxia telangiectasia-like disorder (ATLD) [MIM:604391]. ATLD is a disease with the same clinical feature than ataxia-telangiectasia but with a somewhat milder clinical course.
Sequence similarities

Belongs to the MRE11/RAD32 family.
Post-translational
modifications

Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localization

Nucleus. Localizes to discrete nuclear foci after treatment with genotoxic agents.
Target information above from: UniProt accession P49959 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4361 Human
- Entrez Gene: 17535 Mouse
- Entrez Gene: 64046 Rat
- Omim: 600814 Human
- SwissProt: P49959 Human
- SwissProt: Q61216 Mouse
- SwissProt: Q9JIM0 Rat
- Unigene: 192649 Human
- Unigene: 149071 Mouse
- Unigene: 209040 Rat
see all
Alternative names
- AT like disease antibody
- Ataxia telangiectasia disorder like antibody
- ATLD antibody
- DNA recombination and repair protein antibody
- Double strand break repair protein MRE11A antibody
- Double-strand break repair protein MRE11A antibody
- endo/exonuclease Mre11 antibody
- HNGS1 antibody
- meiotic recombination (S. cerevisiae) 11 homolog A antibody
- Meiotic recombination 11 homolog 1 antibody
- meiotic recombination 11 homolog A (S. cerevisiae) antibody
- Meiotic recombination 11 homolog A antibody
- MmMRE11A antibody
- Mre 11 antibody
- MRE 11a antibody
- MRE 11b antibody
- MRE11 homolog 1 antibody
- MRE11 homolog A antibody
- MRE11 homolog double strand break repair nuclease antibody
- MRE11 meiotic recombination 11 homolog A (S. cerevisiae) antibody
- MRE11 meiotic recombination 11 homolog A antibody
- MRE11_HUMAN antibody
- MRE11A antibody
- MRE11b antibody
- OTTHUMP00000236830 antibody
- OTTHUMP00000236831 antibody
- OTTHUMP00000236832 antibody
- OTTHUMP00000236833 antibody
see all

Western blot - Anti-Mre11 antibody [12D7] (ab214)

Western blot of PC12 (brain)whole cell extracts at 30µg per lane, labeling Mre11 with ab214 at 1:500. A HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
Western blot - Anti-Mre11 antibody [12D7] (ab214)

All lanes : Anti-Mre11 antibody [12D7] (ab214) at 1/1000 dilution

Lane 1 : 293T whole cell extract
Lane 2 : Transfected 293T whole cell extract

Lysates/proteins at 30 µg per lane.

Predicted band size: 79 kDa
Immunohistochemistry (Frozen sections) - Anti-Mre11 antibody [12D7] (ab214)This image is courtesy of an anonymous Abreview

ab214 staining Mre11 in human normal and gastric cancer tissue sections (20µm) by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde for 3 hours, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 23°C. Samples were incubated with primary antibody (1/200 in 3% BSA + 0.1% Triton X-100 in TBS buffer) for 12 hours at 4°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.
See Abreview
Western blot - Anti-Mre11 antibody [12D7] (ab214)This image is courtesy of an anonymous Abreview

All lanes : Anti-Mre11 antibody [12D7] (ab214) at 1/500 dilution

Lane 1 : Mouse breast cancer cell line - whole cell lysate. Transfected with vector control.
Lane 2 : Mouse breast cancer cell line - whole cell lysate. Transfected with knockdown shRNA targeting Mre11 gene.

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP-conjugated Goat anti-mouse IgG polyclonal at 1/1000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 79 kDa
Observed band size: 75 kDa
why is the actual band size different from the predicted?

Exposure time: 1 minute
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-Mre11 antibody [12D7] (ab214)Image supplied by Dr Domenico Delia, Istituto Nazionale Tumori, Italy.

Indirect immunofluorescence to detect localisation of Mre11 in normal lymphoblastoid cells.
(Panel D - negative control).
Western blot - Anti-Mre11 antibody [12D7] (ab214)

This picture was kindly supplied as part of the reviews for ab214 and for ab89 submitted by Anya Polischouk.

The bands represent nuclear (N) and cytoplasmic (C) extract from human lung cancer cells (U1810).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mre11 antibody [12D7] (ab214)This image is courtesy of an anonymous Abreview

ab214 at 1/200 staining normal human bronchus tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in EDTA was peformed. The tissue was blocked in normal rabbit serum and then incubated with the antibody for 1 hour. An HRP conjugated goat polyclonal antibody was used as the secondary.
See Abreview
Flow Cytometry - Anti-Mre11 antibody [12D7] (ab214)

Overlay histogram showing HeLa cells stained with ab214 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab214, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG, H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Roy S et al. p53 orchestrates DNA replication restart homeostasis by suppressing mutagenic RAD52 and POL? pathways. Elife 7:N/A (2018). Read more (PubMed: 29334356) »
Huang D et al. Isoorientin triggers apoptosis of hepatoblastoma by inducing DNA double-strand breaks and suppressing homologous recombination repair. Biomed Pharmacother 101:719-728 (2018). WB ; Human . Read more (PubMed: 29524880) »

See all 47 Publications for this product

Publishing research using ab214? Please let us know so that we can cite the reference in this datasheet.

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Abcam has not validated the combination of species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Pig Cell (LLC-PK1 cell line)

Permeabilization

Yes - 0.5% Triton-X 100

Specification

LLC-PK1 cell line

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: R.T°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 10 2016

Application

Immunohistochemistry (Frozen sections)

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Sample

Human Tissue sections (Normal or gastric cancer tissues)

Specification

Normal or gastric cancer tissues

Permeabilization

Yes - 0.1% Triton X-100

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 20 2014

Application

Immunocytochemistry

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C

Sample

Mouse Cultured Cells (Mouse embryonic fibroblasts)

Specification

Mouse embryonic fibroblasts

Permeabilization

Yes - 0.1% Triton X-100

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 19 2014

Application

Immunocytochemistry

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C

Sample

Human Cultured Cells (Hela cell line)

Specification

Hela cell line

Permeabilization

Yes - 0.1% Triton X-100

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 18 2014

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 23°C

Sample

Human Cell (Human colon cancer cell lines)

Specification

Human colon cancer cell lines

Permeabilization

Yes - 0.3% Triton X-100

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 15 2013

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (7.5)

Sample

Mouse Cell lysate - whole cell (Mouse breast cancer cell line)

Specification

Mouse breast cancer cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Oct 15 2013

Abcam has not validated the combination of species/application used in this Abreview.

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)

Loading amount

15 µg

Specification

Mouse embryonic fibroblasts

Gel Running Conditions

Reduced Denaturing (10% Tris-Gly gel)

Blocking step

LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 08 2011

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - nuclear (human cancer cells (colon, bladder))

Loading amount

20 µg

Specification

human cancer cells (colon, bladder)

Gel Running Conditions

Reduced Denaturing (4-12)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 19°C

Abcam user community

Verified customer

Submitted Apr 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa cells)

Loading amount

20 µg

Specification

HeLa cells

Gel Running Conditions

Reduced Denaturing (10% Tris-Gly gel)

Blocking step

LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 17 2010

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Normal Bronchus)

Specification

Normal Bronchus

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA

Permeabilization

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jun 15 2007

1-10 of 11 Abreviews or Q&A

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Anti-Mre11 antibody [12D7] (ab214)

Overview

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References

This product has been referenced in:

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Mre11 antibody [12D7]

Immunohistochemistry (Frozen sections) abreview for Anti-Mre11 [12D7] antibody

Immunocytochemistry abreview for Anti-Mre11 [12D7] antibody

Immunocytochemistry abreview for Anti-Mre11 [12D7] antibody

Immunocytochemistry/ Immunofluorescence abreview for Anti-Mre11 [12D7] antibody

Western blot abreview for Anti-Mre11 antibody [12D7]

Western blot abreview for Anti-Mre11 antibody [12D7]

Western blot abreview for Anti-Mre11 antibody [12D7]

Western blot abreview for Anti-Mre11 antibody [12D7]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Mre11 antibody [12D7]