Total OXPHOS Blue Native WB Antibody Cocktail (ab110412)


  • Product name
    Total OXPHOS Blue Native WB Antibody Cocktail
  • Assay type
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    Blue Native polyacrylamide gel electrophoresis (BN-PAGE) is a simple and effective way to subfractionate mitochondrial proteins as intact complexes on a single gel (in one dimension). It can be used to detect altered assembly of these complexes arising from mutations in subunits, mutations in assembly factors, or mtDNA depletion. This type of analysis has been performed with biopsy samples, platelets and fibroblast cells from patients with suspected mitochondrial diseases.

    In this method, multisubunit enzymes bind a charged dye Coomassie brilliant blue which allows their electrophoretic separation in the first dimension by the size of the complex. Complexes I-V with masses ranging from 950K to 200K are well resolved in the first dimension. The separated proteins can then be transferred to nitrocellulose membrane/PVDF by electrophoresis and Complexes I, II, III, IV and ATP synthase can be detected by mAbs against CI-NDUFA9 ab14713(MS111), CII-70 kDa subunit ab14715(MS204), CIII-Core protein 2 ab14745(MS304), CIV-subunit IV ab14744(MS407) and CV-alpha subunit ab14748(MS507) respectively. Such one dimensional gels, are best analyzed by using single mAbs against each complex. Other Complex I mAbs are available for BNPAGE, specifically anti-GRIM-19 ab110240(MS103) and anti-20 kDa ab110242(MS105). A sample of purified bovine heart mitochondria ab110338(MS802) is available as a BNPAGE control sample.

    Sometimes a greater separation of enzymes is necessary - it is possible to separate the proteins within each individual complex. To do this, blotting is NOT performed after the first (NATIVE) dimension, instead gels are turned 90 degrees and run in a perpendicular second dimension which is denaturing (NON-NATIVE). In this way the protein subunits within each complex are separated. Abcam provides an optimized pre-mixed cocktail of the mAbs to SIMULTANEOUSLY detect Complexes I-V after 2nd dimension blotting (specifically the cocktail contains ab14713 (MS111), ab14715 (MS204), ab14745 (MS304), ab14744(MS407) and ab14748 (MS507).

    Cocktail Antibodies:

    Mouse monoclonal [20C11B11B11] to (C-I) NDUFA9 (ab14713):
    Amount: 120 µg
    Working Concentration: 2 µg/ml

    Mouse monoclonal [2E3GC12FB2AE2] to (C-II-70) SDHA (ab14715):
    Amount: 6 µg
    Working Concentration: 0.1 µg/ml

    Mouse monoclonal [13G12AF12BB11] to (C-III-Core 2) UQCRC2 (ab14745):
    Amount: 60 µg
    Working Concentration: 1 µg/ml

    Mouse monoclonal [20E8C12] to (C-IV-subunit IV) COX IV(ab14744):
    Amount: 60 µg
    Working Concentration: 1 µg/ml

    Mouse monoclonal [15H4C4 ] to (C-V-alpha) ATP5A (ab14748):
    Amount: 60 µg
    Working Concentration: 1 µg/ml

  • Tested applications
    Suitable for: WBmore details



Our Abpromise guarantee covers the use of ab110412 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 6 µg/ml. The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting


  • Two dimension Blue Native PAGE analysis of fibroblasts that are (A) normal and (B) complex I deficient. It is clear that the complex I deficient cell line shown in B shows no detectable level of complex I. However, all other OXPHOS complexes appear normal including a minor amount of heterocomplex formed between complexes III and IV which is a well documented species. Only a minor mitochondrial enrichment was performed upon these samples cell lines. Each sample represents only 8% of confluent cells taken from a 10 cm diameter tissue culture dish.



    (C-I) NDUFA9 - 36kDa

    (C-II-70) SDHA - 70kDA

    (C-III-Core 2) UQRC2) - 45kDa

    (C-IV subunit IV) COX IV - 15kDa

    (C-V alpha) ATP5A - 55kDa


This product has been referenced in:
  • Gonzalez-Freire M  et al. Skeletal muscle ex vivo mitochondrial respiration parallels decline in vivo oxidative capacity, cardiorespiratory fitness, and muscle strength: The Baltimore Longitudinal Study of Aging. Aging Cell 17:N/A (2018). Read more (PubMed: 29356348) »
  • Bellanti F  et al. Synergistic interaction of fatty acids and oxysterols impairs mitochondrial function and limits liver adaptation during nafld progression. Redox Biol 15:86-96 (2017). WB . Read more (PubMed: 29220698) »

See all 13 Publications for this product

Customer reviews and Q&As

The antibody concentrations in ab110412Total OXPHOS Blue Native WB Antibody Cocktail are only optimized for the example provided on the datasheet.  Maximum signal, minimal background for each protein in cultured cells e.g. fibroblasts.


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Both of these cocktails measure a representative subunit from each of the 5 OXPHOS complexes. Both work on human, mouse and rat.

The MS603 was designed first and is for use when Western blotting a 2D BNPAGE gel to get a good separat...

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Vielen Dank für die Zusammenfassung in Englisch. Ich habe nun eine Antwort von unseren Mitochondrien -Spezialisten aus dem Labor erhalten.

Ich habe die Antwort nicht übersetzt um Übersetzungsfehler zu vermeiden:


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Thank you for contacting us.

In theory, any antibody can be used in the “second” dimension of the Blue Native electrophoresis. In the first dimension, proteins are in their native form and only antibodies validated to work in this...

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Thank you for contacting us. As you may have seen, there is an image of a 2D denatured gel on the product datasheet. Unfortunately, we have an image of a denatured gel not available, and we have not used this product for blotting the 1D native gel b...

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Thank you for contacting us. The Blue Native Western Blot does not require specific gels. Native acrylamide gels can be poured by hand. While it is possible to use a single acrylamide concentration we recommend the use of a linear acrylamide co...

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Thank you for your call today and for your questions about ab110412 (MS603). I have found that this antibody is suitable for use after 2-D separation. We do have some data at the bottom of the datasheet that shows staining following 2-D separat...

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