Overview

  • Product name
    Anti-NDUFS3 antibody [3F9DD2]
    See all NDUFS3 primary antibodies
  • Description
    Mouse monoclonal [3F9DD2] to NDUFS3
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Fr, IHC-Glut, Flow Cyt, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Zebrafish
  • Immunogen

    Whole mitochondria from Human Heart.

  • Positive control
    • Isolated mitochondria from Human, Bovine Rat and Mouse hearts; Cultured Human fibroblasts; HeLa cells; Human colon and cerebellum tissues.
  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab110246 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
IHC-Fr Use at an assay dependent concentration. PubMed: 25223649
IHC-Glut 1/100.
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 5 - 10 µg/ml.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB Use a concentration of 0.5 µg/ml. Predicted molecular weight: 30 kDa.

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 [3F9DD2] (ab110246) 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 monoclonal HRP conjugate at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 30 kDa
    Additional bands at: 260 kDa (possible non-specific binding), 40 kDa (possible non-specific binding)


    Exposure time: 1 minute

    See Abreview

  • All lanes : Anti-NDUFS3 antibody [3F9DD2] (ab110246) at 0.5 µg/ml

    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

    Predicted band size: 30 kDa

  • ab110246, at 10 µg/ml, staining NDUFS3 in cultured Human fibroblasts by Immunofluorescence. The cells were fixed, treated for heat-induced antigen retrieval, permeabilized and then labeled with ab110246, followed by a fluorescent goat-anti-mouse IgG.
  • ab110246, at 1 µg/ml, staining NDUFS3 in HeLa cells by Flow Cytometry (Blue).

    Isotype control antibody, at 1 µg/ml, staining (red).

  • ab110246, at 1/100, staining NDUFS3 in Human colon tissue fixed with 4% PFA by Immunohistochemistry.

    ab110246 was labeled with a universal probe (which recognizes antibodies raised in both mouse and rabbit species) for 30 minutes and a HRP-polymer probe for 30 minutes. ab110246 was diluted and incubated for 1 hour with the target. Antigen retrieval was not required.

  • ab110246, at 1/1000, staining NDUFS3 in formalin fixed and paraffin embedded Human cerebellum tissue by Immunohistochemistry.

    ab110246 was labeled with a universal probe (which recognizes antibodies raised in both mouse and rabbit species) for 30 minutes and a HRP-polymer probe for 30 minutes. ab110246 was diluted and incubated with the target for 1 hour. Antigen retrieval performed by 1 min pressure cooking with 1mmol EDTA, pH8.

  • ab110246 staining NDUFS3 in Human colon tissue sections from a normal ageing patient by Immunohistochemistry. Kindly provided by Dr. L. Greaves and D. Turnbull, Mitochondrial Research Group, Newcastle University. For more details, see Taylor et al., J. Clin. Invest. 112:1351-1360 (2003).

References

This product has been referenced in:
  • Lee YL  et al. Epigallocatechin-3-gallate alleviates bladder overactivity in a rat model with metabolic syndrome and ovarian hormone deficiency through mitochondria apoptosis pathways. Sci Rep 8:5358 (2018). Read more (PubMed: 29599473) »
  • Shi B  et al. SNAPIN is critical for lysosomal acidification and autophagosome maturation in macrophages. Autophagy 13:285-301 (2017). Read more (PubMed: 27929705) »
See all 45 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (SH-SY5Y)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
SH-SY5Y
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Mr. Chun Chau SUNG

Verified customer

Submitted Aug 07 2018

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

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 10 2015

Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Total protein in input
30 µg

Abcam user community

Verified customer

Submitted Jan 07 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (HeLa cells)
Specification
HeLa cells
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 24 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient Tris-Gly gel)
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Blocking step
LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 19 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient Tris-Gly gel)
Sample
Mouse Cell lysate - whole cell (MEFs)
Specification
MEFs
Blocking step
LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 19 2014

Answer

Thank you for your reply.
For a use on tissue sections I can recommend to use an antibody directed against NDUFS3 (or Complex I 30kDa subunit) such as ab110246. The link to the datasheet of this antibody is www.abcam.com/ab110246.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you very much for contacting us with your question.
The lab does use an antigen retrieval step while staining cultured cells with ab110246, so I would recommend using this step. Our ICC protocol with antigen retrieval can be found at this link-
https://www.abcam.com/index.html?pageconfig=resource&rid=14733
I would suggest either fixing with ice-cold acetone for 1-10 minutes, which will also permeabilize the cells, or fixing with 4% PFA and permeabilizing with 100 uM digitonin or 0.5% saponin which are milder than Triton and better for membrane-associated antigens.
I hope that this information will be helpful, but if you have any further questions or need anything else please let me know.

Read More

Answer

Thank you for contacting us.

That’s right we immunized the whole complex. This protein is very immunoreactive, we have generated many monoclonals against this subunit.

Subunits are large compared to the epitope of an antibody binding site so it is quite easy to get lots of antibodies against specific proteins from a large complex.

The trick is to identify exactly which subunits they bind specifically to and in this case that was done in the way I mentioned. This is definitely against mitochondrial NDSUFS3, we have good data and a strong publication record for this clone.

If there are bands in the cytosol these could be mitochondrial contaminants of the cytosol or its possible NDUFS3 protein in transit to the mitochondrion.

We don’t see an extra band with this antibody as you can see from our datasheet blot. We don’t see a 70 kDa band (though there is a 75 kDa protein in the complex (NDUFS1) we don’t see any cross reactive). Likely an extra band is coming from non specific binding which may be cured by changing the blocking conditions.

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

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1-10 of 12 Abreviews or Q&A

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