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Validated using a knockout cell line

Anti-p95/NBS1 antibody [7E4A2] (ab181729)

Submit a review Submit a question References (2)
Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Immunocytochemistry/ Immunofluorescence - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
  • Flow Cytometry - Anti-p95/NBS1 antibody [7E4A2] (ab181729)

Key features and details

  • Mouse monoclonal [7E4A2] to p95/NBS1
  • Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
  • Knockout validated
  • Reacts with: Rat, Human
  • Isotype: IgG2a

Overview

  • Product name

    Anti-p95/NBS1 antibody [7E4A2]
    See all p95/NBS1 primary antibodies

  • Description

    Mouse monoclonal [7E4A2] to p95/NBS1

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Orangutan

  • Immunogen

    Recombinant fragment corresponding to Human p95/NBS1 aa 467-615. Expressed in E. Coli.
    Sequence:

    ERDEENQEMSSCKSARIETSCSLLEQTQPATPSLWKNKEQHLSENEPVDT NSDNNLFTDTDLKSIVKNSASKSHAAEKLRSNKKREMDDVAIEDEVLEQL FKDTKPELEIDVKVQKQEEDVNVRKRPRMDIETNDTFSDEAVPESSKIS


    Database link: O60934
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • A549, Jurkat and PC-12 cell lysates; p95/NBS1 (aa 467-615) recombinant protein; p95/NBS1 (aa 467-615)-hIgGFc transfected HEK293 cell lysate; Hela cells; Human cervical cancer and Human rectum cancer tissues. WB: A431 cell lysate.

  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR150721-8 are from Tissue Culture Supernatant.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181729 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
1/200 - 1/400.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P
1/200 - 1/1000.
WB
1/500 - 1/2000. Predicted molecular weight: 84 kDa.
ICC/IF
1/200 - 1/1000.
Notes
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.
WB
1/500 - 1/2000. Predicted molecular weight: 84 kDa.
ICC/IF
1/200 - 1/1000.

Target

  • 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
    modifications

    Phosphorylated 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

  • Database links

    see all

  • Alternative names

    • AT V1 antibody
    • AT V2 antibody
    • ATV antibody
    • Cell cycle regulatory protein p95 antibody
    • FLJ10155 antibody
    • MGC87362 antibody
    • Nbn antibody
    • NBN_HUMAN antibody
    • NBS 1 antibody
    • NBS antibody
    • NBS1 antibody
    • Nibrin antibody
    • Nijmegen breakage syndrome 1 (nibrin) antibody
    • Nijmegen breakage syndrome antibody
    • Nijmegen breakage syndrome protein 1 antibody
    • p95 antibody
    • p95 protein of the MRE11/RAD50 complex antibody
    see all

Images

  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    All lanes : Anti-p95/NBS1 antibody [7E4A2] (ab181729) 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: 84 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 [7E4A2] 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, ab181729 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.

  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    All lanes : Anti-p95/NBS1 antibody [7E4A2] (ab181729) at 1/500 dilution

    Lane 1 : A549 cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : PC-12 cell lysate

    Predicted band size: 84 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Immunocytochemistry/ Immunofluorescence - Anti-p95/NBS1 antibody [7E4A2] (ab181729)

    Immunofluorescent analysis of HeLa cells labeling p95/NBS1 with ab181729 at 1/200 (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.

  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    All lanes : Anti-p95/NBS1 antibody [7E4A2] (ab181729) at 1/500 dilution

    Lane 1 : non-transfected HEK293 cell lysate
    Lane 2 : p95/NBS1 (aa 467-615)-hIgGFc transfected HEK293 cell lysate

    Predicted band size: 84 kDa

  • Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Western blot - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Anti-p95/NBS1 antibody [7E4A2] (ab181729) at 1/500 dilution + p95/NBS1 (aa 467-615) human recombinant protein

    Predicted band size: 84 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)

    Immunohistochemical analysis of paraffin embedded Human rectum cancer tissue labeling p95/NBS1 with ab181729 at 1/200 with DAB staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [7E4A2] (ab181729)

    Immunohistochemical analysis of paraffin embedded Human cervical cancer tissue labeling p95/NBS1 with ab181729 at 1/200 with DAB staining.

  • Flow Cytometry - Anti-p95/NBS1 antibody [7E4A2] (ab181729)
    Flow Cytometry - Anti-p95/NBS1 antibody [7E4A2] (ab181729)

    Flow Cytometrical analysis of HeLa cells labeling p95/NBS1 with ab181729 at 1/200 (green) compared to a negative control antibody (red).

References (2)

Publishing research using ab181729? Please let us know so that we can cite the reference in this datasheet.

ab181729 has been referenced in 2 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
  • Ye Z  et al. GRB2 enforces homology-directed repair initiation by MRE11. Sci Adv 7:N/A (2021). PubMed: 34348893

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