存放说明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.
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纯化说明Purified from ascites.
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.
|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).|
|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.|
功能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.
疾病相关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.
序列相似性Belongs to the MRE11/RAD32 family.
翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
细胞定位Nucleus. Localizes to discrete nuclear foci after treatment with genotoxic agents.
- Information by UniProt
- AT like disease antibody
- Ataxia telangiectasia disorder like antibody
- ATLD antibody
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.
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
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.
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.
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
Indirect immunofluorescence to detect localisation of Mre11 in normal lymphoblastoid cells.
(Panel D - negative control).
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).
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.
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.
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) »