The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/500 - 1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 84 kDa).
1/50 - 1/100.
Is unsuitable for Flow Cyt or IHC-P.
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.
Ubiquitous. Expressed at high levels in testis.
Nijmegen breakage syndrome Breast cancer Aplastic anemia Defects in NBN might play a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).
Contains 1 BRCT domain. Contains 1 FHA 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.
Phosphorylated by ATM in response of ionizing radiation, and such phosphorylation is responsible intra-S phase checkpoint control and telomere maintenance.
Nucleus. Nucleus, PML body. Chromosome, telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
Western blot - Anti-p95 NBS1 (phospho S343) antibody [EP178] (ab109453)
All lanes : Anti-p95 NBS1 (phospho S343) antibody [EP178] (ab109453) at 1/5000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma) cells whole cell lysates Lane 2 : HeLa (human cervix adenocarcinoma) cells were treated with Etopside whole cell lysates Lane 3 : HeLa (human 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
Dot Blot - Anti-p95 NBS1 (phospho S343) antibody [EP178] (ab109453)
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.