RabMAb

Anti-p95 NBS1抗体[Y112] - ChIP Grade (ab32074)

概述

  • 产品名称Anti-p95 NBS1抗体[Y112] - ChIP Grade
    参阅全部 p95 NBS1 一抗
  • 描述
    兔单克隆抗体[Y112] to p95 NBS1 - ChIP Grade
  • 经测试应用适用于: WB, IP, IHC-P, ICC/IF, Flow Cyt, ChIP/Chipmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human p95 NBS1 aa 650-750 (C terminal).

  • 阳性对照
    • WB: HeLa and Jurkat cell lysates. IHC-P: Human testis and skin carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

性能

应用

Our Abpromise guarantee covers the use of ab32074 in the following tested applications.

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

应用 Ab评论 说明
WB 1/1000 - 1/10000. Predicted molecular weight: 85 kDa.

For unpurified use at 1/1000 - 1/2000.

IP 1/60.
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/50.

ICC/IF 1/100.

For unpurified use at 1/50.

Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ChIP/Chip Use at an assay dependent concentration. PubMed: 21255732

靶标

  • 功能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.
  • 组织特异性Ubiquitous. Expressed at high levels in testis.
  • 疾病相关Nijmegen breakage syndrome
    Breast cancer
    Aplastic anemia
    Defects in NBN might play a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).
  • 序列相似性Contains 1 BRCT domain.
    Contains 1 FHA domain.
  • 结构域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.
  • 翻译后修饰Phosphorylated by ATM in response of ionizing radiation, and such phosphorylation is responsible intra-S phase checkpoint control and telomere maintenance.
  • 细胞定位Nucleus. Nucleus, PML body. Chromosome, telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 antibody [Y112] - ChIP Grade 图像

  • All lanes : Anti-p95 NBS1 antibody [Y112] - ChIP Grade (ab32074) at 1/1200 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 85 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling p95 NBS1 with purified ab32074 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

  • Immunocytochemsitry/Immunofluorescence analysis of HeLa cells labelling p95 NBS1 (green) with unpurified ab32074 at 1/20. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • Overlay histogram showing HeLa cells stained with unpurified ab32074 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32074, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • All lanes : Anti-p95 NBS1 antibody [Y112] - ChIP Grade (ab32074) at 1/10000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 85 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling p95 NBS1 with unpurified ab32074 at 1/20. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with hematoxylin.

  • Immunocytochemsitry/Immunofluorescence analysis of HeLa cells labelling p95 NBS1 (green) with purified ab32074 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • Unpurified ab32074 staining p95 NBS1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were treated +/- 5 Gy IR. Cells were fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 1 hour. Samples were incubated with primary antibody (1/500 in 5% BSA) for 1 hour. A FITC-conjugated donkey anti-rabbit IgG polyclonal (1/150) was used as the secondary antibody.

    See Abreview

Anti-p95 NBS1 antibody [Y112] - ChIP Grade (ab32074)参考文献

This product has been referenced in:
  • Khair L  et al. Nbs1 ChIP-Seq Identifies Off-Target DNA Double-Strand Breaks Induced by AID in Activated Splenic B Cells. PLoS Genet 11:e1005438 (2015). ChIP ; Mouse . Read more (PubMed: 26263206) »
  • Yousif AS  et al. Differential regulation of S-region hypermutation and class-switch recombination by noncanonical functions of uracil DNA glycosylase. Proc Natl Acad Sci U S A 111:E1016-24 (2014). Read more (PubMed: 24591630) »

See all 11 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 50 µg
Specification HeLa
Treatment 10 Gy IR
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Apr 24 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - Triton X-100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 May 30 2014

Thank you for your inquiry.

I am happy to confirm that the concentration of lot GR4228-7 is 0.645 mg/mL.

I also would like to comment on the seemingly low concentration:

The IgG produced by rabbits have greater affinity an...

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The concentration of ab32074 lot GR4228-5is : 0.645 mg/mL.

Thank you for your call today and for keeping me updated about this situation.

I am sending the three vials thatwe havein stock. These vials will be arriving on the free of charge order 1060035, and I expect them to arrive tomorrow.

Read More

Thank you for your enquiry.

I can confirm that p95 NBS1 antibody [Y112] (ab32074) was tested onsquamous cell carcinoma of the skin. This would be a suitable positive control.

I hope this information will be helpful to you. If you ...

Read More

Thank you for contacting Abcam.



The concentration for ab32074, lot #GR50908-2 is *****mg/mL.



Please let me know if there is anything else I can help you with.

Thanks for your call today and for letting us know about the trouble with ab32074 in ChIP.

As we discussed, I am sending a free of charge vial of ab32074 from lot GR50908-2 on the order ***, which should arrive tomorrow.

I've looked...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human carotid plaque tissue)
Specification Human carotid plaque tissue
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Jurkat cells)
Loading amount 100000 cells
Specification Jurkat cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 19 2010

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