重组Anti-p95/NBS1抗体[Y112] (ab32074)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y112] to p95/NBS1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-p95/NBS1抗体[Y112]
参阅全部 p95/NBS1 一抗 -
描述
兔单克隆抗体[Y112] to p95/NBS1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-Pmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Human
预测可用于: Mouse, Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, Jurkat and Wild-type A431 cell lysates. IHC-P: Human testis and skin carcinoma tissues.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y112 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32074于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (3) |
1/1000 - 1/10000. Predicted molecular weight: 85 kDa.
For unpurified use at 1/1000 - 1/2000. |
IHC-P | (1) |
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/50. |
说明 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 85 kDa. For unpurified use at 1/1000 - 1/2000. |
IHC-P
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/50. |
靶标
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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
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数据库链接
- Entrez Gene: 4683 Human
- Entrez Gene: 27354 Mouse
- Entrez Gene: 85482 Rat
- Omim: 602667 Human
- SwissProt: O60934 Human
- SwissProt: Q9R207 Mouse
- SwissProt: Q9JIL9 Rat
- Unigene: 492208 Human
see all -
别名
- AT V1 antibody
- AT V2 antibody
- ATV antibody
see all
图片
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/1000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : NBN knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab32074 observed at 90 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab32074 was shown to react with p95/NBS1 in wild-type A431 cells in Western blot with loss of signal observed in NBN knockout cell line ab269506 (NBN knockout cell lysate ab269668). Wild-type A431 and NBN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab32074 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NBN knockout HeLa cell lysate
Lane 3 : Wild-type HeLa nuclear cell lysate
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?Lanes 1-3: Merged signal (red and green). Green - ab32074 observed at 95 kDa.
ab32074 Anti-p95/NBS1 antibody [Y112] was shown to specifically react with p95/NBS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261834 (knockout cell lysate ab257111) was used. Wild-type and p95/NBS1 knockout samples were subjected to SDS-PAGE. ab32074, Anti-GAPDH antibody [6C5] - Cytoplasmic Loading Control (ab8245) and Anti-Histone H3 (ab176842) - Nuclear Loading Control were incubated overnight at 4°C at 1 in 1000 dilution, 1 in 20000 dilution and 1 in 1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773), Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/1200 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] (ab32074)
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.
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All lanes : Anti-p95/NBS1 antibody [Y112] (ab32074) at 1/10000 dilution (purified)
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p95/NBS1 antibody [Y112] (ab32074)
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.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (26)
ab32074 被引用在 26 文献中.
- Zhang B et al. NBS1 is required for SPO11-linked DNA double-strand break repair in male meiosis. Cell Death Differ 27:2176-2190 (2020). PubMed: 31965061
- Dellino GI et al. Release of paused RNA polymerase II at specific loci favors DNA double-strand-break formation and promotes cancer translocations. Nat Genet 51:1011-1023 (2019). PubMed: 31110352
- Leimbacher PA et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell 74:571-583.e8 (2019). PubMed: 30898438
- Carvajal-Maldonado D et al. Perturbing cohesin dynamics drives MRE11 nuclease-dependent replication fork slowing. Nucleic Acids Res 47:1294-1310 (2019). PubMed: 29917110
- Afzal S et al. Rat Glioma Cell-Based Functional Characterization of Anti-Stress and Protein Deaggregation Activities in the Marine Carotenoids, Astaxanthin and Fucoxanthin. Mar Drugs 17:N/A (2019). PubMed: 30909572