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使用敲除细胞株进行验证重组RabMAb

重组Anti-NDUFS3抗体[EPR12782] - C-terminal (ab177471)

Reviews (2) Submit a question References (8)
Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Flow Cytometry (Intracellular) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
  • Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12782] to NDUFS3 - C-terminal
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-NDUFS3抗体[EPR12782] - C-terminal
    参阅全部 NDUFS3 一抗

  • 描述

    兔单克隆抗体[EPR12782] to NDUFS3 - C-terminal

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details

  • 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.

  • 阳性对照

    • HepG2, 293T, HL60 and fetal heart lysates. Human heart and kidney tissue. 293 cells.

  • 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
    For more information see here.

    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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab177471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
Flow Cyt (Intra)
1/200.
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 30 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/50 - 1/100.
IP
1/10 - 1/100.
说明
Flow Cyt (Intra)
1/200.
WB
1/1000 - 1/10000. Predicted molecular weight: 30 kDa.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/100.
IP
1/10 - 1/100.

靶标

  • 功能

    Core subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I) that is believed to belong to the minimal assembly required for catalysis. Complex I functions in the transfer of electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone.

  • 序列相似性

    Belongs to the complex I 30 kDa subunit family.

  • 细胞定位

    Mitochondrion inner membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

      • CI 30 antibody
      • CI 30KD antibody
      • CI-30kD antibody
      • Complex I 30KD antibody
      • Complex I 30kDa subunit antibody
      • COMPLEX I, MITOCHONDRIAL RESPIRATORY CHAIN, 30-KD SUBUNIT antibody
      • Complex I-30kD antibody
      • mitochondrial antibody
      • NADH coenzyme Q reductase antibody
      • NADH dehydrogenase (ubiquinone) Fe S protein 3 30kDa antibody
      • NADH dehydrogenase [ubiquinone] iron sulfur protein 3 mitochondrial antibody
      • NADH dehydrogenase [ubiquinone] iron-sulfur protein 3 antibody
      • NADH dehydrogenase ubiquinone 30 kDa subunit antibody
      • NADH-ubiquinone oxidoreductase 30 kDa subunit antibody
      • NADH-Ubiquinone Oxidoreductase Fe-S Protein 3 antibody
      • NDUFS3 antibody
      • NDUS3_HUMAN antibody
      see all

    图片

    • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : NDUFS3 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 30 kDa
      Observed band size: 27 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab177471 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab177471 was shown to react with NDUFS3 in HEK-293T wild-type cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab177471 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

      Immunofluorescence analysis of 293 cells labeling NDUFS3 with ab177471 at 1/50 dilution  (green). DAPI nuclear staining (blue).

    • Flow Cytometry (Intracellular) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Flow Cytometry (Intracellular) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

      Intracellular Flow Cytometry analysis of HEK293 (Human embryonic kidney epithelial cell) cells labeling NDUFS3 (red) with purified ab177471 at a 1/200 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue). 

       

       

    • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)Image is courtesy of an anonymous Abreview.
      All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

      Lanes 1-2 : Whole cell lysates from NDUFS3 KO cells.
      Lanes 3-4 : Whole cell lysates from NDUFS3 WT cells.
      Lanes 5-6 : Isolated mitochondria from NDUFS3 KO Cells.
      Lanes 7-8 : Isolated mitochondria from NDUFS3 WT Cells.

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit polyclonal HRP conjugate. at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 30 kDa


      Exposure time: 1 minute

      See Abreview

    • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : NDUFS3 knockout HEK-293T cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : HL60 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 30 kDa
      Observed band size: 30 kDa



      Lanes 1- 4: Merged signal (red and green). Green - ab177471 observed at 30 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab177471 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type HEK-293T and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab177471 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

      Western blot analysis on immunoprecipitation pellet from (1) human fetal heart lysate or (2) 1XPBS (negative control) using ab177471at 1/10 dilution, and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

      Immunohistochemcal analysis of paraffin-embedded Human heart tissue labeling NDUFS3 with ab177471 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

      Immunohistochemcal analysis of paraffin-embedded Human kidney tissue labeling NDUFS3 with ab177471 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

      Lane 1 : HepG2 lysate
      Lane 2 : 293T lysate
      Lane 3 : HL60 lysate
      Lane 4 : Fetal heart lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

      Developed using the ECL technique.

      Predicted band size: 30 kDa

    • Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)
      Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

    文献 (8)

    发表研究结果有使用 ab177471？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab177471 被引用在 8 文献中.

    • Balderas E  et al. Mitochondrial calcium uniporter stabilization preserves energetic homeostasis during Complex I impairment. Nat Commun 13:2769 (2022). PubMed: 35589699
    • Zhou C  et al. CDC25C is a prognostic biomarker and correlated with mitochondrial homeostasis in pancreatic adenocarcinoma. Bioengineered 13:13089-13107 (2022). PubMed: 35615982
    • Zeng X  et al. Inhibition of mitochondrial and cytosolic calpain attenuates atrophy in myotubes co-cultured with colon carcinoma cells. Oncol Lett 21:124 (2021). PubMed: 33552245
    • Johnstone T  et al. Biallelic variants in two complex I genes cause abnormal splicing defects in probands with mild Leigh syndrome. Mol Genet Metab 131:98-106 (2020). PubMed: 33097395
    • Kory N  et al. MCART1/SLC25A51 is required for mitochondrial NAD transport. Sci Adv 6:N/A (2020). PubMed: 33087354
    • Simon MT  et al. Novel mutations in the mitochondrial complex I assembly gene NDUFAF5 reveal heterogeneous phenotypes. Mol Genet Metab 126:53-63 (2019). PubMed: 30473481
    • Kurelac I  et al. Inducing cancer indolence by targeting mitochondrial Complex I is potentiated by blocking macrophage-mediated adaptive responses. Nat Commun 10:903 (2019). PubMed: 30796225
    • Chen XF  et al. Effect of puerarin in promoting fatty acid oxidation by increasing mitochondrial oxidative capacity and biogenesis in skeletal muscle in diabetic rats. Nutr Diabetes 8:1 (2018). PubMed: 29330446

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    1-2 of 2 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-NDUFS3 antibody [EPR12782] - C-terminal

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK293)
    Permeabilization
    Yes - 0.5% Triton X-100
    Specification
    HEK293
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Neutral buffered formalin
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 May 20 2019

    Western blot abreview for Anti-NDUFS3 antibody [EPR12782] - C-terminal

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - other (osteosarcoma)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    50 µg
    Specification
    osteosarcoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Luisa Iommarini

    Verified customer

    提交于 Nov 03 2015

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