Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NDUFS3 antibody [17D95] (ab14711)

Key features and details

  • Mouse monoclonal [17D95] to NDUFS3
  • Suitable for: WB, Flow Cyt
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG2a

Overview

  • Product name

    Anti-NDUFS3 antibody [17D95]
    See all NDUFS3 primary antibodies

  • Description

    Mouse monoclonal [17D95] to NDUFS3

  • Host species

    Mouse

  • Tested Applications & Species

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

  • Immunogen

    Tissue, cells or virus. This information is considered to be commercially sensitive.

  • Positive control

    • Human heart mitochondria. In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HepG2 cells.

  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)
    Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)
    All lanes : Anti-NDUFS3 antibody [17D95] (ab14711) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human heart tissue lysate - total protein (ab29431)
    Lane 3 : Heart (Mouse) Tissue Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 30 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 57 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute


    The band observed at 26 kDa could potentially be a cleaved form of NDUFS3 due to the presence of a 36 amino acid transit peptide.
  • Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)
    Western blot - Anti-NDUFS3 antibody [17D95] (ab14711) Image is courtesy of an anonymous AbReview
    All lanes : Anti-NDUFS3 antibody [17D95] (ab14711) 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-mouse polyclonal HRP conjugate at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 30 kDa
    Additional bands at: 22 kDa (possible non-specific binding), 40 kDa (possible non-specific binding)


    Exposure time: 1 minute

    See Abreview

  • Flow Cytometry - Anti-NDUFS3 antibody [17D95] (ab14711)
    Flow Cytometry - Anti-NDUFS3 antibody [17D95] (ab14711)
    Overlay histogram showing HepG2 cells stained with ab14711 (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 (ab14711, 1µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)
    Western blot - Anti-NDUFS3 antibody [17D95] (ab14711)
    All lanes : Anti-NDUFS3 antibody [17D95] (ab14711)

    Lane 1 : Isolated mitochondria from Human heart at 5 µg
    Lane 2 : Isolated mitochondria from Bovine heart at 4 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: 30 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.

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