Anti-p95 NBS1 (phospho S432) 抗体 (ab12297)

製品の概要

  • 製品名Anti-p95 NBS1 (phospho S432) antibody
    p95 NBS1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to p95 NBS1 (phospho S432)
  • 特異性This antibody detects the immunising phospho-peptide but not the non-phospho peptide on dot blots. By western blot on NBS1 transfected ILB1 cells it detects a band representing NBS1 on wt NBS1 transfected cells, but not on S432A NBS1 transfected cells. This confirms that the antibody is specific for phospho S432 NBS1.
  • アプリケーション適用あり: WBmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human p95 NBS1, phosphorylated at S432.

    (Peptide available as ab12312.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: ILB1 cells transfected with wild type NBS1.
  • 特記事項


    This antibody recognises a highly conserved novel potential CDK phosphorylation site in NBS1. There is already a significant body of data suggesting that mammalian NBS1 and the Mre11/Rad50/Nbs1 (MRN) complex may be cell cycle regulated, for instance, the activity of the yeast MRN complex (Mre11/Rad50/Xrs2) is regulated in a cell cycle-dependent manner through phosphorylation by cyclin-dependent kinase 1 (CDK1), however, the phosphorylation sites in yeast have not been mapped. It has been shown using mass spectrometry that this residue is phosphorylated in vivo in human NBS1. This antibody confirms the existence of that modification.

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファーPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • 一次抗体 備考This antibody recognises a highly conserved novel potential CDK phosphorylation site in NBS1. There is already a significant body of data suggesting that mammalian NBS1 and the Mre11/Rad50/Nbs1 (MRN) complex may be cell cycle regulated, for instance, the activity of the yeast MRN complex (Mre11/Rad50/Xrs2) is regulated in a cell cycle-dependent manner through phosphorylation by cyclin-dependent kinase 1 (CDK1), however, the phosphorylation sites in yeast have not been mapped. It has been shown using mass spectrometry that this residue is phosphorylated in vivo in human NBS1. This antibody confirms the existence of that modification.
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab12297 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use at an assay dependent dilution. Predicted molecular weight: 85 kDa. Optimal dilutions/concentrations should be determined by the end user. Can be blocked with p95 NBS1 phospho S432 peptide. This antibody performs better when used subsequent to IP with an NBS1 antibody (see pictures below

ターゲット情報

  • 機能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.
  • 組織特異性Ubiquitous. Expressed at high levels in testis.
  • 関連疾患Nijmegen breakage syndrome
    Breast cancer
    Aplastic anemia
    Defects in NBN might play a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).
  • 配列類似性Contains 1 BRCT domain.
    Contains 1 FHA domain.
  • ドメイン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.
  • 翻訳後修飾Phosphorylated by ATM in response of ionizing radiation, and such phosphorylation is responsible intra-S phase checkpoint control and telomere maintenance.
  • 細胞内局在Nucleus. Nucleus, PML body. Chromosome, telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-p95 NBS1 (phospho S432) antibody 画像

  • Dot blot using ab12297 on phospho and non-phospho peptides corresponding to the immunising peptide sequence.

    Dots 1: 1ng of peptide
    Dots 2: 10ng of peptide
    Dots 3: 100ng of peptide

  • Lanes 1 - 2 : Anti-p95 NBS1 (phospho S432) antibody (ab12297)
    Lanes 3 - 4 : NBS1 antibody (recognizes modified and unmodified NBS1)

    Lane 1 : IP'd lysate from cells transfected with wild type (WT) NBS1.
    Lane 2 : IP'd lysate from cells transfected with mutant (S432A) NBS1.
    Lane 3 : IP'd lysate from cells transfected with wild type (WT) NBS1.
    Lane 4 : IP'd lysate from cells transfected with mutant (S432A) NBS1.


    Performed under reducing conditions.

    Predicted band size : 85 kDa
    Observed band size : 85 kDa

    Western blot on the IP fraction from ILB1 cells that have been IP'd with an NBS1 antibody. ILB1 cells (NBS1 deficient) were transfected to either express WT NBS1 or S432A NBS1.

    Lane 1: ab12297 used on IP fraction of WT NBS1 expressors.
    Lane 2: ab12297 used on IP fraction of S432A NBS1 expressors.
    Lane 3: NBS1 antibody used  on IP fraction of WT NBS1 expressors.
    Lane 4: NBS1 antibody used on IP fraction of S432A NBS1 expressors.

    Western blot on the IP fraction from ILB1 cells that have been IP'd with an NBS1 antibody that recognizes both the phosphorylated and non-phosphorylated NBS1. ILB1 cells (NBS1 deficient) were transfected to either express WT NBS1 or S432A NBS1.
  • All lanes : Anti-p95 NBS1 (phospho S432) antibody (ab12297)

    Lane 1 : ILB1 cells transfected with empty vector.
    Lane 2 : ILB1 cells transfected with wild type NBS1.
    Lane 3 : ILB1 cells transfected with mutant (S432A) NBS1.


    Performed under reducing conditions.

    Predicted band size : 85 kDa

    Western blot using ab12297 on transfected ILB1 cells (NBS1 deficient).

    Lane 1: ILB1 cells + empty vector
    Lane 2: ILB1 cells + WT NBS1
    Lane 3: ILB1 cells + S432A NBS1

    The band representing NBS1 is indicated by an arrow.

    Western blot using ab12297 on transfected ILB1 cells (NBS1 deficient). The band representing NBS1 is indicated by an arrow.
  • Anti-p95 NBS1 (phospho S432) antibody (ab12297) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

    Secondary
    Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 85 kDa
    Observed band size : 98,100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 150 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab12297 overnight at 4°C. Antibody binding was detected using an anti-rabbit IgG VHH single domain antibody conjugated to HRP, and visualised using ECL development solution ab133406.

Anti-p95 NBS1 (phospho S432) antibody (ab12297) 使用論文

ab12297 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-X100 in PBS
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Dr. Kirk McManus

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投稿 Feb 10 2012

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