Overview

  • Product name
    Anti-NDUFS3 antibody [17D95]
    See all NDUFS3 primary antibodies
  • Description
    Mouse monoclonal [17D95] to NDUFS3
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish
  • Immunogen

    Purified cow mitochondrial complex I.

  • 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.

Applications

Our Abpromise guarantee covers the use of ab14711 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
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 30 kDa).
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the complex I 30 kDa subunit family.
  • Cellular localization
    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • 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.
  • 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

  • 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.

  • 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.

References

This product has been referenced in:
  • Baertling F  et al. NDUFA9 point mutations cause a variable mitochondrial complex I assembly defect. Clin Genet 93:111-118 (2018). WB ; Human . Read more (PubMed: 28671271) »
  • Pareek G  et al. Loss of the Drosophila m-AAA mitochondrial protease paraplegin results in mitochondrial dysfunction, shortened lifespan, and neuronal and muscular degeneration. Cell Death Dis 9:304 (2018). Read more (PubMed: 29467464) »
See all 58 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Nov 03 2015

Question
Answer

Thank you for contacting us. It was nice talking to you.

As discussed on phone I am happy to provide you 5% discount for purchase of any one of the discussed product + free ab123069
Will be happy to provide 4, 100% Abreview discount codes; using these you will get 4 free antibodies. Please click the link for more info

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

Alternatively, I can provide you 4, 100% Image discounts and guarantee of replacement or refund if the antibody does not work. You only need to submit an Abreview with Images and protocol details after using these antibodies.

Once you submit Abreview we will be able to give you Abpoints ˜220 per antibody so you can earn 880 abpoints that can be used further to get money off from your next order. (100GBP off with 700 Abpoints)

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you very much for your patient. I really appreciate the time taken to send us further details and images. The reactivity of this antibody with Caenorhabditis elegans has not been tested in the lab. However, there are specific publications confirming the reactivity of the antibody against this species: Yang YY et al. The effect of different ubiquinones on lifespan in Caenorhabditis elegans. Mech Ageing Dev 130:370-6 (2009). WB; Caenorhabditis elegans. PubMed: 19428456 In any case, in the future I would encourage you to use ab14711 for targeting NDUFS3 in WB with Caenorhabditis elegans, as we have lab data to support it, and I will happy to offer you a free of charge replacement of ab14711, or a credit note in compensation. I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

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