Anti-NDUFS3 antibody [3F9DD2] (ab110246)

Reviews (7)Q&A (5)References (49)

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
    see all
  • 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

  • Western blot - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Western blot - Anti-NDUFS3 antibody [3F9DD2] (ab110246)This image is courtesy of an anonymous AbReview
    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

  • Western blot - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Western blot - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    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.
  • Flow Cytometry - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Flow Cytometry - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    ab110246, at 1 µg/ml, staining NDUFS3 in HeLa cells by Flow Cytometry (Blue).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [3F9DD2] (ab110246)
    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:
  • Thomas LW  et al. CHCHD4 confers metabolic vulnerabilities to tumour cells through its control of the mitochondrial respiratory chain. Cancer Metab 7:2 (2019). Read more (PubMed: 30886710) »
  • Gopal RK  et al. Early loss of mitochondrial complex I and rewiring of glutathione metabolism in renal oncocytoma. Proc Natl Acad Sci U S A 115:E6283-E6290 (2018). Read more (PubMed: 29915083) »
See all 49 Publications for this product

Publishing research using ab110246? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 12 Abreviews or Q&A

Western blot abreview for Anti-NDUFS3 antibody [3F9DD2]

 Average
Abreviews
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
Read More

Mr. Chun Chau SUNG

Verified customer

Submitted Aug 07 2018

Western blot abreview for Anti-NDUFS3 antibody [3F9DD2]

 Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Nov 03 2015

Western blot abreview for Anti-NDUFS3 antibody [3F9DD2]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 10 2015

Immunoprecipitation abreview for Anti-NDUFS3 antibody [3F9DD2]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jan 07 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-NDUFS3 antibody [3F9DD2]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Nov 24 2014

Western blot abreview for Anti-NDUFS3 antibody [3F9DD2]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Sep 19 2014

Western blot abreview for Anti-NDUFS3 antibody [3F9DD2]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Sep 19 2014

Question

Actually, these products do not respond to my request. As written below, we would need to measure mitochondrial activity not in tissue or cell extracts, but in histological sections.
Do you have anything like that?
Thanks,

Read More
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

Question

Inquiry: Hello, We purchased ab110246 for immunocytochemistry staining of mitochondrial complex I on cultured cells. However, with standard 4% paraformaldehyde, subsequent 0.1% TX100 permeabilization, and 10 ug/mL ab110246, we were unable to detect an immunofluorescence signal. It was briefly mentioned on your product sheet that the fibroblasts were fixed, treated for heat-induced antigen retrieval, and permeabilized. What are your specific conditions for fixation, and permeabilization, and is antigen retrieval necessary for this antibody? Many thanks.

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

Question

if I understand correctly, you used the entire the complex I to produce this Ab. Sinceraly I can't understand how this Ab could be specific, for only one of the 45 complex I subunits. Apart of this our problenm is that when we test the Ab on cytosol we noticed a band corresponding to the NDUFS3 full-lenght protein and other upper MW bands (approximately 70 KDa). Do you have any ideas of what they could be?

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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