Recombinant Anti-p95/NBS1 (phospho S343) antibody [EP178] (ab109453)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP178] to p95/NBS1 (phospho S343)
- Suitable for: ICC/IF, Dot blot, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-p95/NBS1 (phospho S343) antibody [EP178]
See all p95/NBS1 primary antibodies -
Description
Rabbit monoclonal [EP178] to p95/NBS1 (phospho S343) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Dot blot, WBmore details
Unsuitable for: Flow Cyt or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, HeLa cell lysates (treated and untreated with Etopside). ICC/IF: Jurkat cells treated with Etoposide (25uM) for 2 h
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP178 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109453 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 |
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ICC/IF |
1/50 - 1/100.
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Dot blot |
Use at an assay dependent concentration.
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WB |
1/500 - 1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 84 kDa).
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Notes |
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ICC/IF
1/50 - 1/100. |
Dot blot
Use at an assay dependent concentration. |
WB
1/500 - 1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 84 kDa). |
Target
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Function
Component of the MRE11-RAD50-NBN (MRN complex) which plays a critical role in the cellular response to DNA damage and the maintenance of chromosome integrity. The complex is involved in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity, cell cycle checkpoint control 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. NBN modulate the DNA damage signal sensing by recruiting PI3/PI4-kinase family members ATM, ATR, and probably DNA-PKcs to the DNA damage sites and activating their functions. It can also recruit MRE11 and RAD50 to the proximity of DSBs by an interaction with the histone H2AX. NBN also functions in telomere length maintenance by generating the 3' overhang which serves as a primer for telomerase dependent telomere elongation. NBN is a major player in the control of intra-S-phase checkpoint and there is some evidence that NBN is involved in G1 and G2 checkpoints. The roles of NBS1/MRN encompass DNA damage sensor, signal transducer, and effector, which enable cells to maintain DNA integrity and genomic stability. Forms a complex with RBBP8 to link DNA double-strand break sensing to resection. Enhances AKT1 phosphorylation possibly by association with the mTORC2 complex. -
Tissue specificity
Ubiquitous. Expressed at high levels in testis. -
Involvement in disease
Nijmegen breakage syndrome
Breast cancer
Aplastic anemia
Defects in NBN might play a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL). -
Sequence similarities
Contains 1 BRCT domain.
Contains 1 FHA domain. -
Domain
The FHA and BRCT domains are likely to have a crucial role for both binding to histone H2AFX and for relocalization of MRE11/RAD50 complex to the vicinity of DNA damage.
The C-terminal domain contains a MRE11-binding site, and this interaction is required for the nuclear localization of the MRN complex.
The EEXXXDDL motif at the C-terminus is required for the interaction with ATM and its recruitment to sites of DNA damage and promote the phosphorylation of ATM substrates, leading to the events of DNA damage response. -
Post-translational
modificationsPhosphorylated by ATM in response of ionizing radiation, and such phosphorylation is responsible intra-S phase checkpoint control and telomere maintenance. -
Cellular localization
Nucleus. Nucleus, PML body. Chromosome, telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents. - Information by UniProt
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Database links
- Entrez Gene: 4683 Human
- Omim: 602667 Human
- SwissProt: O60934 Human
- Unigene: 492208 Human
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Alternative names
- AT V1 antibody
- AT V2 antibody
- ATV antibody
see all
Images
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All lanes : Anti-p95/NBS1 (phospho S343) antibody [EP178] (ab109453) at 1/5000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cells whole cell lysates
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were treated with Etopside whole cell lysates
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were treated with Etopside whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 84 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking and diluting buffer: 5% NFDM/TBST
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Immunocytochemistry analysis of Jurkat (human T cell leukemia T lymphocyte) labeling p95/NBS1 with purified ab109453 at 1/100 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.27 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody.Confocal image showing increased nuclear staining in Jurkat cells treated with Etoposide (25uM) for 2 h.
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Dot blot analysis of p95/NBS1 (pS343) peptide (Lane 1) and p95/NBS1 non-phospho peptide (Lane 2) labelling p95/NBS1 (phospho S343) with ab109453 at a dilution of 1/1000. A Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab109453 has been referenced in 3 publications.
- Tang D et al. MYC/NBS1-Mediated DNA Damage Response is Involved in the Inhibitory Effect of Hydroxysafflor Yellow A on Glioma Cells. Drug Des Devel Ther 15:1749-1763 (2021). PubMed: 33953544
- Tang Z et al. Active DNA end processing in micronuclei of ovarian cancer cells. BMC Cancer 18:426 (2018). PubMed: 29661159
- Schoenherr RM et al. Commercially available antibodies can be applied in quantitative multiplexed peptide immunoaffinity enrichment targeted mass spectrometry assays. Proteomics 16:2141-5 (2016). PubMed: 27094115