Overview

  • Product nameAnti-p95 NBS1 antibodySee all p95 NBS1 primary antibodies ...
  • Description
    Rabbit polyclonal to p95 NBS1
  • Tested applicationsWB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Human p95 NBS1.

    (Peptide available as ab24289.)

  • Positive control
    • ab23996 gave a positive result in the following lysates: MRC5 whole cell lysate (SV40 transformed immortal (WT) fibroblasts), Hela whole cell, NIH3T3 whole cell, HepG2 whole cell and Jurkat whole cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab23996 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB 1/1000 - 1/5000. Detects a band of approximately 95 kDa (predicted molecular weight: 85 kDa).Can be blocked with Human p95 NBS1 peptide (ab24289). Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionComponent 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.
  • Tissue specificityUbiquitous. Expressed at high levels in testis.
  • Involvement in diseaseNijmegen breakage syndrome (NBS) [MIM:251260]: A disorder characterized by chromosomal instability, radiation sensitivity, microcephaly, growth retardation, immunodeficiency and predisposition to cancer, particularly to lymphoid malignancies. Note=The disease is caused by mutations affecting the gene represented in this entry.
    Breast cancer (BC) [MIM:114480]: A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.
    Aplastic anemia (AA) [MIM:609135]: A form of anemia in which the bone marrow fails to produce adequate numbers of peripheral blood elements. It is characterized by peripheral pancytopenia and marrow hypoplasia. Note=Disease susceptibility may be associated with variations affecting the gene represented in this entry.
    Note=Defects in NBN might play a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).
  • Sequence similaritiesContains 1 BRCT domain.
    Contains 1 FHA domain.
  • DomainThe 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 localizationNucleus. Chromosome > telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
  • Target information above from: UniProt accession O60934 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • AT V1 antibody
    • AT V2 antibody
    • ATV antibody
    • ATV antibody
    • Cell cycle regulatory protein p95 antibody
    • FLJ10155 antibody
    • MGC87362 antibody
    • NBN antibody
    • NBN_HUMAN antibody
    • NBS 1 antibody
    • NBS antibody
    • NBS antibody
    • NBS1 antibody
    • Nibrin antibody
    • Nibrin antibody
    • Nijmegen breakage syndrome 1 (nibrin) antibody
    • Nijmegen breakage syndrome antibody
    • Nijmegen breakage syndrome protein 1 antibody
    • p95 antibody
    • p95 antibody
    • p95 protein of the MRE11/RAD50 complex antibody
    see all

Anti-p95 NBS1 antibody images

  • All lanes : Anti-p95 NBS1 antibody (ab23996) at 1/5000 dilution

    Lane 1 : Lysate from cells engineered to be NBS1 defective (SV40 transformed, immortal fibroblasts)
    Lane 2 : MRC5 cell lysate (SV40 transformed immortal (WT) fibroblasts)

    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?)
  • All lanes : Anti-p95 NBS1 antibody (ab23996) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 85 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 125 kDa,55 kDa,65 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab23996 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23996, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 and MCF7 cells at 1µg/ml.

References for Anti-p95 NBS1 antibody (ab23996)

This product has been referenced in:
  • Fan S  et al. DIM (3,3'-diindolylmethane) confers protection against ionizing radiation by a unique mechanism. Proc Natl Acad Sci U S A 110:18650-5 (2013). WB ; Rat . Read more (PubMed: 24127581) »
  • Jamil S  et al. MCL-1 localizes to sites of DNA damage and regulates DNA damage response. Cell Cycle 9:2843-55 (2010). Read more (PubMed: 20647761) »

See all 2 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Nov 30 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"