- Datasheet
- Specific References (30)
- Protocols
Overview
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Product nameAnti-Rad50 antibody [13B3/2C6]
See all Rad50 primary antibodies -
DescriptionMouse monoclonal [13B3/2C6] to Rad50
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Host speciesMouse
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Tested applicationsSuitable for: Flow Cyt, IHC-P, ICC/IF, WB, IPmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
Fusion protein containing the complete coding region (amino acids 1-425) of RAD50 expressed in E.coli.
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Positive control
- Raji cell lysate HEK293 cell lysate
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General notesClone 2C6 recognizes Rad50, a 153kDa protein involved in DNA double strand break repair. Rad50 is associated with Mre11 and p95 (Nibrin) to form a multiprotein complex involved in the double strand break repair process.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityMonoclonal
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Clone number13B3/2C6
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MyelomaNS1
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IsotypeIgG1
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Light chain typekappa
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Research areas
Associated products
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab89 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 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| IHC-P | 1/400. | |
| ICC/IF | 1/200. See Abreview. | |
| WB | 1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 146 kDa). | |
| IP | Use at an assay dependent concentration. |
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
- Omim: 604040 Human
- SwissProt: Q92878 Human
- SwissProt: P70388 Mouse
- Unigene: 633509 Human
- Unigene: 4888 Mouse
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Alternative names
- DNA repair protein RAD50 antibody
- hRAD50 antibody
- hsRAD50 antibody
see all
Images
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![Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)](https://www.abcam.com/ps/products/0/ab89/Images/ab2141.jpg)
Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)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).
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).
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![Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)](https://www.abcam.com/ps/products/0/ab89/Images/ab89_1.jpg)
Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)Anti-Rad50 antibody [13B3/2C6] (ab89) at 1/1000 dilution + 50 ug Human HEK293 total cell lysate
Secondary
HRP conjugated Donkey Anti-Mouse IgG
Developed using the ECL technique.
Performed under non-reducing conditions.
Predicted band size: 146 kDa
Observed band size: 160 kDa (why is the actual band size different from the predicted?)
Additional bands at: 300 kDa (possible non-specific binding)
Exposure time: 30 seconds
This image is courtesy of an Abreview submitted by Philippe Szankasi on 24 February 2006. -
![Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [13B3/2C6] (ab89)](https://www.abcam.com/ps/products/0/ab89/Images/Rad50-Primary-antibodies-ab89-1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [13B3/2C6] (ab89)Image from Kadaja M et. al., PLoS Pathog. 2009 April; 5(4): e1000397 (Fig 4).ab89 staining Rad50 in HPV transfected Hela cells by Immunocytochemistry/ Immunofluorescence. Cells were washed twice with phosphate-buffered saline (PBS), permeabilized with 0.5% Triton X-100 in CSK buffer (10 mM Hepes-KOH, pH 7.4; 300 mM sucrose; 100 mM NaCl; 3 mM KCl) for 2 min on ice, and fixed with 4% paraformaldehyde. Fixed cells were treated with 0.5% Triton X-100 in PBS, followed by 3 PBS washes for 5 min each at RT. After blocking with 3% bovine serum albumin (BSA) in PBS at RT for 30 min, the cells were incubated with primary antibody in antibody-binding solution (3% BSA in PBS) at RT for 30 min. An Alexa Fluor® 488 conjugated anti mouse secondary was used.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad50 antibody [13B3/2C6] (ab89)](https://www.abcam.com/ps/products/0/ab89/Images/Rad50-Primary-antibodies-ab89-2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad50 antibody [13B3/2C6] (ab89)ab89 (1µg/ml) staining RAD50 in human placenta, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
![Flow Cytometry - Anti-Rad50 antibody [13B3/2C6] (ab89)](https://www.abcam.com/ps/products/0/ab89/Images/Rad50-Primary-antibodies-ab89-3.jpg)
Flow Cytometry - Anti-Rad50 antibody [13B3/2C6] (ab89)Overlay histogram showing HeLa cells stained with ab89 (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 (ab89, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse 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.
Protocols
References
This product has been referenced in:
- Luo M et al. Unrepaired DNA damage in macrophages causes elevation of particulate matter- induced airway inflammatory response. Aging (Albany NY) 10:549-560 (2018). Read more (PubMed: 29661984) »
- Brandt S et al. Lack of MRE11-RAD50-NBS1 (MRN) complex detection occurs frequently in low-grade epithelial ovarian cancer. BMC Cancer 17:44 (2017). Read more (PubMed: 28073364) »
Customer reviews and Q&As
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