Recombinant Anti-Rad50 antibody [EPR3466(2)] (ab124682)
- Datasheet
- References (1)
- Protocols
Overview
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Product nameAnti-Rad50 antibody [EPR3466(2)]
See all Rad50 primary antibodies -
DescriptionRabbit monoclonal [EPR3466(2)] to Rad50
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Host speciesRabbit
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Tested applicationsSuitable for: Flow Cyt, ICC/IF, WBmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide within Human Rad50 aa 1250-1350. The exact sequence is proprietary.
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Positive control
- K562, MOLT4, Jurkat and MCF7 cell lysates.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
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Storage bufferpH: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR3466(2)
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab124682 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 | 1/80. | |
| ICC/IF | 1/100. | |
| WB | 1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 154 kDa). |
Target
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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.
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Tissue specificityExpressed at very low level in most tissues, except in testis where it is expressed at higher level. Expressed in fibroblasts.
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Involvement in diseaseDefects in RAD50 are the cause of Nijmegen breakage syndrome-like disorder (NBSLD) [MIM:613078]; also called NBS-like disorder or RAD50 deficiency. NBSLD is a disorder similar to Nijmegen breakage syndrome and characterized by chromosomal instability, radiation sensitivity, microcephaly, growth retardation, short stature and bird-like face. Immunodeficiency is absent.
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Sequence similaritiesBelongs to the SMC family. RAD50 subfamily.
Contains 1 zinc-hook domain. -
DomainThe zinc-hook, which separates the large intramolecular coiled coil regions, contains 2 Cys residues that coordinate one molecule of zinc with the help of the 2 Cys residues of the zinc-hook of another RAD50 molecule, thereby forming a V-shaped homodimer. The two heads of the homodimer, which constitute the ATP-binding domain, interact with the MRE11A homodimer.
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Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localizationNucleus. Chromosome > telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
- Information by UniProt
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Database links
- Entrez Gene: 10111 Human
- Entrez Gene: 19360 Mouse
- Entrez Gene: 64012 Rat
- Omim: 604040 Human
- SwissProt: Q92878 Human
- SwissProt: P70388 Mouse
- SwissProt: Q9JIL8 Rat
- Unigene: 633509 Human
see all -
Alternative names
- DNA repair protein RAD50 antibody
- hRAD50 antibody
- hsRAD50 antibody
see all
Images
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![Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-296770-ab124682WB2.jpg)
Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/5000 dilution (purified)
Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
Lane 2 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 154 kDaBlocking and diluting buffer: 5% NFDM/TBST
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![Flow Cytometry - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-7-ab124682FC1.jpg)
Flow Cytometry - Anti-Rad50 antibody [EPR3466(2)] (ab124682)Flow Cytometry analysis of U-2 OS (Human bone osteosarcoma epithelial cell) cells labeling Rad50 with purified ab124682 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-6-ab124682ICCIF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Rad50 with Purified ab124682 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 . ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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![Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-296769-ab124682WB1.jpg)
Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (purified)
Lane 1 : Human fetal brain lysates
Lane 2 : Mouse thymus lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 154 kDaBlocking and diluting buffer: 5% NFDM/TBST
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![Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-296771-ab124682WB3.jpg)
Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (purified) + Rat testis lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 154 kDaBlocking and diluting buffer: 5% NFDM/TBST
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![Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/ab124682-236498-124682-IF.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)This image is courtesy of an anonymous AbreviewUnpurified ab124682 staining Rad50 in the U2OS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 5% BSA for 60 minutes. Samples were incubated with undiluted primary antibody for 2 hours. An undiluted FITC conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
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![Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)](https://www.abcam.com/ps/products/124/ab124682/Images/Rad50-Primary-antibodies-ab124682-2.jpg)
Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (unpurified)
Lane 1 : K562 cell lysate
Lane 2 : MOLT4 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 154 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Datasheets and documents
References
This product has been referenced in:
- Chen Y et al. EGFR tyrosine kinase inhibitor HS-10182 increases radiation sensitivity in non-small cell lung cancers with EGFR T790M mutation. Cancer Biol Med 15:39-51 (2018). Read more (PubMed: 29545967) »
