The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1-2µg for 106 cells.
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).
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 79 kDa (predicted molecular weight: 79 kDa). (see Robinson et al).
Use a concentration of 1 - 5 µg/ml. (for normal lymphoblastoid cell lines).
Use at an assay dependent concentration.
FunctionComponent 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 diseaseDefects 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 similaritiesBelongs to the MRE11/RAD32 family.
Post-translational modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus. Localizes to discrete nuclear foci after treatment with genotoxic agents.
Double strand break repair protein MRE11A antibody
Double-strand break repair protein MRE11A antibody
endo/exonuclease Mre11 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
Mre 11 antibody
MRE 11a antibody
MRE 11b antibody
MRE11 homolog 1 antibody
MRE11 homolog A antibody
MRE11 meiotic recombination 11 homolog A (S. cerevisiae) antibody
MRE11 meiotic recombination 11 homolog A antibody
Anti-Mre11 antibody [12D7] images
Immunohistochemistry (Frozen sections) - Anti-Mre11 [12D7] antibody (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.
Western blot - Anti-Mre11 [12D7] antibody (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 HRP-conjugated Goat anti-mouse IgG polyclonal at 1/1000 dilution Developed using the ECL technique
Immunocytochemistry/ Immunofluorescence - 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)
Predicted band size : 79 kDa
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) - 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.
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.