Anti-p95/NBS1 antibody [7E4C2] (ab181780)
Key features and details
- Mouse monoclonal [7E4C2] to p95/NBS1
- Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-p95/NBS1 antibody [7E4C2]
See all p95/NBS1 primary antibodies -
Description
Mouse monoclonal [7E4C2] to p95/NBS1 -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment corresponding to Human p95/NBS1 aa 467-615. Expressed in E. Coli.
Sequence:ERDEENQEMSSCKSARIETSCSLLEQTQPATPSLWKNKEQHLSENEPVDT NSDNNLFTDTDLKSIVKNSASKSHAAEKLRSNKKREMDDVAIEDEVLEQL FKDTKPELEIDVKVQKQEEDVNVRKRPRMDIETNDTFSDEAVPESSKIS
Database link: O60934 -
Positive control
- Human p95/NBS1 recombinant protein; HEK293 cell lysate, transfected with p95/NBS1 (amino acids 467-615)-IgGFc; Jurkat cell lysate; HeLa cells; Human cervical cancer and colon cancer tissues. WB: A431 cell lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 99% PBS -
Concentration information loading...
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Purity
Protein G purified -
Purification notes
Purified from tissue culture supernatant. -
Clonality
Monoclonal -
Clone number
7E4C2 -
Isotype
IgG2a -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181780 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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WB |
1/500 - 1/2000. Predicted molecular weight: 85 kDa.
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ICC/IF |
1/200 - 1/1000.
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Flow Cyt |
1/200 - 1/400.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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IHC-P |
1/200 - 1/1000.
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Notes |
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WB
1/500 - 1/2000. Predicted molecular weight: 85 kDa. |
ICC/IF
1/200 - 1/1000. |
Flow Cyt
1/200 - 1/400. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
IHC-P
1/200 - 1/1000. |
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 antibody [7E4C2] (ab181780) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : NBN knockout A549 cell lysate
Lane 3 : Wild-type A431 cell lysate
Lane 4 : NBN knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Anti-NBN antibody [7E4C2] (ab181780) staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab181780 was shown to bind specifically to NBN. A band was observed at 95 kDa in wild-type A549 cell lysates with no signal observed at this size in NBN knockout cell line. To generate this image, wild-type and NBN knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-p95/NBS1 antibody [7E4C2] (ab181780) at 1/500 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : NBN knockout A431 cell lysate
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-p95/NBS1 antibody [7E4C2] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab181780 was shown to bind specifically to p95/NBS1. A band was observed at 95 kDa in wild-type A431 cell lysates with no signal observed at this size in NBN knockout cell line ab269506 (knockout cell lysate ab269668). To generate this image, wild-type and NBN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Anti-p95/NBS1 antibody [7E4C2] (ab181780) at 1/500 dilution + Human p95/NBS1 recombinant protein (amino acids 467-615)
Predicted band size: 85 kDa -
All lanes : Anti-p95/NBS1 antibody [7E4C2] (ab181780) at 1/500 dilution
Lane 1 : HEK293 cell lysate, non-transfected
Lane 2 : HEK293 cell lysate, transfected with p95/NBS1 (amino acids 467-615)-hIgGFc
Predicted band size: 85 kDa -
Anti-p95/NBS1 antibody [7E4C2] (ab181780) at 1/500 dilution + Jurkat cell lysate
Predicted band size: 85 kDa -
Immunofluorescent analysis of HeLa cells labeling p95/NBS1 with ab181780 at 1/200 dilution (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.
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Flow cytometric analysis of HeLa cells labeling p95/NBS1 with ab181780 at 1/200 dilution (green); negative control (red).
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Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling p95/NBS1 with ab181780 at 1/200 dilution followed by DAB staining.
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Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling p95/NBS1 with ab181780 at 1/200 dilution followed by DAB staining.
Protocols
Datasheets and documents
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Datasheet download
References (0)
ab181780 has not yet been referenced specifically in any publications.