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Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

Key features and details

  • Mouse monoclonal [20C11B11B11] to NDUFA9
  • Suitable for: IHC-P, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-NDUFA9 antibody [20C11B11B11]
    See all NDUFA9 primary antibodies

  • Description

    Mouse monoclonal [20C11B11B11] to NDUFA9

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    Tissue, cells or virus corresponding to Cow NDUFA9.

  • Positive control

    • WB: WI38 and NIH 3T3 whole cell lysates, human testis tissue lysate and human, cow, rat and mouse heart mitochondria. IHC-P: Human spinal column tissue. Flow Cyt: HepG2 cells.

  • General notes

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to NDUFA9 has been knockout validated in Western blot. The expected band for NDUFA9 was observed in wild type cells and the band was not seen in knockout cells.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Images

  • Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: NDUFA9 knockout HAP1 cell lysate (20 µg)
    Lane 3: WI38 cell lysate (20 µg)
    Lane 4: NIH3T3 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab14713 observed at 40 kDa. Red - loading control, ab176560, observed at 52 kDa.

    ab14713 was shown to specifically react with NDUFA9 in wild-type HAP1 cells. No band was observed when NDUFA9 knockout HAP1 samples were used. Wild-type and NDUFA9 knockout samples were subjected to SDS-PAGE. ab14713 and ab176560 (loading control to alpha tubulin) were diluted at 1μg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    ab14713 a 1/100 dilution staining NDUFA9 in Human spinal column tissue by Immunohistochemistry (Formalin/PFA-Fixed paraffin-embedded sections). Antibody was incubated with the sample for 1 hour. Sections were incubated in peroxidase-conjugated rabbit anti-mouse secondary (diluted 1/100 in 4% BSA in PBST) for 1 hour at room temperature. Sections were washed x3 in PBST and peroxidase activity was demonstrated using kit. Antigen retrieval was performed by 1 minute of pressure cooking with 1 mmol EDTA pH 8.0.
  • Flow Cytometry - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    Flow Cytometry - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

    Overlay histogram showing HepG2 cells stained with ab14713 (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 (ab14713, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    All lanes : Anti-NDUFA9 antibody [20C11B11B11] (ab14713) at 1 µg/ml

    Lane 1 : WI38 (Human lung fibroblast cell line) Whole Cell Lysate
    Lane 2 : Human testis tissue lysate - total protein (ab30257)
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 40 kDa
    Observed band size: 36 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    The band observed at 36 kDa could potentially be a cleaved form of NDUFA9 due to the presence of a 35 amino acid transit peptide.
  • Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    Western blot - Anti-NDUFA9 antibody [20C11B11B11] (ab14713)
    All lanes : Anti-NDUFA9 antibody [20C11B11B11] (ab14713)

    Lane 1 : Isolated mitochondria from Human heart at 5 µg
    Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
    Lane 3 : Isolated mitochondria from Rat heart at 10 µg
    Lane 4 : Isolated mitochondria from Mouse Heart at 10 µg

    Secondary
    All lanes : Goat anti-Mouse IgG

    Predicted band size: 40 kDa
    Observed band size: 37 kDa why is the actual band size different from the predicted?

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