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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-NDUFB8 antibody [EPR15961] (ab192878)

Reviews (1) Submit a question References (6)
Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Flow Cytometry (Intracellular) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Immunoprecipitation - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Immunocytochemistry/ Immunofluorescence - Anti-NDUFB8 antibody [EPR15961] (ab192878)
  • Anti-NDUFB8 antibody [EPR15961] (ab192878)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15961] to NDUFB8
  • Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-NDUFB8 antibody [EPR15961]
    See all NDUFB8 primary antibodies

  • Description

    Rabbit monoclonal [EPR15961] to NDUFB8

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Human fetal liver, fetal heart and tonsil lysates; HeLa, C6, PC12, NIH 3T3 and RAW 264.7 cell lysates; Human kidney, Mouse brain and Rat kidney tissues; HeLa cells.

  • General notes

    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.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab192878 in the following tested applications.

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

Application Abreviews Notes
ICC/IF
1/50.
IP
1/20.
WB (1)
1/1000 - 1/10000. Detects a band of approximately 19 kDa (predicted molecular weight: 22 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
ICC/IF
1/50.
IP
1/20.
WB
1/1000 - 1/10000. Detects a band of approximately 19 kDa (predicted molecular weight: 22 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)
Use at an assay dependent concentration.

Target

  • Function

    Accessory subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I), that is believed not to be involved in 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.

  • Sequence similarities

    Belongs to the complex I NDUFB8 subunit family.

  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • ASHI antibody
      • CI-ASHI antibody
      • Complex I ASHI subunit antibody
      • Complex I-ASHI antibody
      • mitochondrial antibody
      • NADH dehydrogenase (ubiquinone) 1 beta subcomplex 8 19kDa antibody
      • NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8 antibody
      • NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8 mitochondrial antibody
      • NADH-ubiquinone oxidoreductase ASHI subunit antibody
      • NDUB8_HUMAN antibody
      • NDUFB8 antibody
      see all

    Images

    • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      All lanes : Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : NDUFB8 knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 22 kDa
      Observed band size: 19 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab192878 observed at 19 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab192878 was shown to specifically react with NDUFB8 in wild-type HAP1 cells as signal was lost in NDUFB8 knockout cells. Wild-type and NDUFB8 knockout samples were subjected to SDS-PAGE. Ab192878 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Flow Cytometry (Intracellular) - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling NDUFB8 antibody (red) with purified ab192878 at a dilution of 1/30. Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody. 

       

       

    • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      All lanes : Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/5000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Human tonsil lysate
      Lane 3 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 22 kDa
      Observed band size: 19 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/20000 dilution + Human fetal heart lysate at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 22 kDa
      Observed band size: 19 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Western blot - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      All lanes : Anti-NDUFB8 antibody [EPR15961] (ab192878) at 1/5000 dilution

      Lane 1 : C6 cell lysate
      Lane 2 : RAW 264.7 cell lysate
      Lane 3 : PC-12 cell lysate
      Lane 4 : NIH/3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

      Predicted band size: 22 kDa
      Observed band size: 19 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling NDUFB8 with ab192878 at 1/500 dilution followed by pre-diluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Immunoprecipitation - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Western blot analysis of NDUFB8 immunoprecipitated from Human fetal heart lysate using ab192878 at 1/20 dilution. Lane 1: Human fetal liver lysate. Lane 2: PBS instead of Human fetal liver lysate.
      Secondary: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Immunocytochemistry/ Immunofluorescence - Anti-NDUFB8 antibody [EPR15961] (ab192878)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized HeLa cells labeling NDUFB8 with ab192878 at 1/50 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) (ab150077) secondary antibody at 1/400 dilution. Nuclear counter stained is DAPI (blue).

    • Anti-NDUFB8 antibody [EPR15961] (ab192878)
      Anti-NDUFB8 antibody [EPR15961] (ab192878)

    References (6)

    Publishing research using ab192878? Please let us know so that we can cite the reference in this datasheet.

    ab192878 has been referenced in 6 publications.

    • Bandara AB  et al. Complex II subunit SDHD is critical for cell growth and metabolism, which can be partially restored with a synthetic ubiquinone analog. BMC Mol Cell Biol 22:35 (2021). PubMed: 34118887
    • Zhao W  et al. Estrogen Deficiency Induces Mitochondrial Damage Prior to Emergence of Cognitive Deficits in a Postmenopausal Mouse Model. Front Aging Neurosci 13:713819 (2021). PubMed: 34335235
    • Song Y  et al. Screening of Potential Biomarkers for Gastric Cancer with Diagnostic Value Using Label-free Global Proteome Analysis. Genomics Proteomics Bioinformatics 18:679-695 (2020). PubMed: 33607292
    • Liu Z  et al. IFI6 depletion inhibits esophageal squamous cell carcinoma progression through reactive oxygen species accumulation via mitochondrial dysfunction and endoplasmic reticulum stress. J Exp Clin Cancer Res 39:144 (2020). PubMed: 32727517
    • Lee JS  et al. Targeting Oxidative Phosphorylation Reverses Drug Resistance in Cancer Cells by Blocking Autophagy Recycling. Cells 9:N/A (2020). PubMed: 32883024
    • Wang Y  et al. The STAT-ROS cycle extends IFN-induced cancer cell apoptosis. Int J Oncol 52:305-313 (2018). PubMed: 29115415

    Customer reviews and Q&As

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    Western blot abreview for Anti-NDUFB8 antibody [EPR15961]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T, HCT116)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    35 µg
    Specification
    HEK293T, HCT116
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°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.

    Abcam user community

    Verified customer

    Submitted Aug 31 2019

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