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Total OXPHOS Rodent WB Antibody Cocktail (ab110413)

Reviews (17)Q&A (26)References (842)
Western blot - MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail (ab110413)
  • AbReview
  • CI subunit NDUFB8 (ab110242)
  • CII-30kDa (ab14714)
  • CIII-Core protein 2 (ab14745)
  • CIV subunit I (ab14705)
  • CV alpha subunit (ab14748)
  • CV alpha subunit (ab14748)

Key features and details

  • Assay type: Quantitative

Overview

  • Product name

    Total OXPHOS Rodent WB Antibody Cocktail

  • Assay type

    Quantitative

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human, Cynomolgus monkey

  • Product overview

    Total OXPHOS Rodent WB Antibody Cocktail ab110413 is an optimized cocktail of high quality antibodies for analyzing relative levels of OXPHOS complexes in rat or mouse mitochondria by western blot. 


    This OXPHOS cocktail contains 5 mouse mAbs, one each against CI subunit NDUFB8 (ab110242), CII-30kDa (ab14714), CIII-Core protein 2 (ab14745), CIV subunit I (ab14705) and CV alpha subunit (ab14748) as an optimized premixed cocktail. The kit is suitable for Western Blotting analysis of the relative levels of the 5 OXPHOS complexes in mitochondrial preparations from mouse, rat, human, or bovine sources. The positive control supplied with the cocktail is ab110341.


    Altered levels of assembly can arise from mutations in individual subunits, mutations in assembly factors for the complex(es), mtDNA depletion or as a result of physiological and or pathological changes e.g. hormone treatment, exercise, diet or oxidative stress.


    The mAbs in the cocktail were chosen because they are each against a subunit that is labile when its complex is not assembled. Also, the different subunits are easily resolved in SDS-PAGE (see protocols).


    Note: Mouse tissue samples can easily be contaminated with antibodies from the animal's blood. To avoid background bands, use ab110413 with an anti-mouse secondary against native antibodies. Also, COXI is a highly hydrophobic protein and appears as a broad band at ~35 kDa (not at its true molecular weight at 57 kDa). It is very sensitive to heating. Therefore, the samples, including the positive control, should not be heated over 50°C before loaded on the gel.


    The Western blot cocktail is supplied at a concentration of 1.5 mg/mL.


    We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot.


    Store the antibody cocktail at 4°C and the control sample at -80°C.

  • Notes

    Related products

    Review the mitochondrial assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

  • Tested applications

    Suitable for: WBmore details

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab110413 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 at an assay dependent concentration.

The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting.

We recommend using PBS with 1% milk as the antibody diluent.

Store the antibody cocktail at 4°C and the control sample at -80°C.

We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot. Please contact us for more details.
Notes
WB
Use at an assay dependent concentration.

The antibody cocktail (1.5 mg/mL) should be diluted 250x to a final working concentration of 6.0 µg/mL for Western blotting.

We recommend using PBS with 1% milk as the antibody diluent.

Store the antibody cocktail at 4°C and the control sample at -80°C.

We recommend using a high pH CAPS / PVDF transfer protocol when using this antibody for Western blot. Please contact us for more details.

Images

  • Western blot - MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail (ab110413)
    Western blot - MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail (ab110413)

    Rat liver mitochondria labeled with ab110413 (MS604). The sample in lane 1 was kept at room temperature, whereas the remaining three samples were heated to 37°C, 50°C, and 100°C, respectively. This blot shows that boiling of samples leads to a decrease in signal due to aggregation of proteins therefore heating samples at or close to boiling is not recommended.

  • AbReview
    AbReviewThis image is courtesy of Aida Adlimoghaddam

    Isolated mitochondria from mice brain (control and Alzheimer’s disease (AD)) labeled with ab110413 at 1/1000 dilution in 5% BSA.

    This image shows NADH dehydrogenase beta subcomplex subunit 8 of Complex I (NDUFB8), succinate dehydrogenase subunit B of Complex II (SDHB), cytochrome c oxidase subunit 1 of Complex IV (MTCO1), cytochrome b-c1 complex subunit 2 of Complex III (UQCRC2) and ATP synthase subunit alpha of Complex V (ATP5A).

    See Abreview

  • CI subunit NDUFB8 (ab110242)
    CI subunit NDUFB8 (ab110242)

    Isolated mitochondria from heart of human (5 µg - Lane 1), cow (1 ug - Lane 2), rat (10 µg - Lane 3), mouse (10 µg - Lane 4) labeling CI subunit NDUFB8 with ab110242 at 0.5 ug/ml. 

    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.

  • CII-30kDa (ab14714)
    CII-30kDa (ab14714)

    Isolated mitochondria from heart of human (5 µg - Lane 1), cow (1 ug - Lane 2), rat (10 µg - Lane 3), mouse (10 µg - Lane 4), HepG2 (human liver hepatocellular carcinoma cell line) whole cell  lysate (20 µg -Lane 5) labeling CII-30kDa with ab14714 at 5 µg/mL. Secondary antibody is a goat anti-mouse antibody.

    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.

  • CIII-Core protein 2 (ab14745)
    CIII-Core protein 2 (ab14745)

    Human skeletal muscle tissue lysate, (10 µg - Lane 1) and Ramos (human Burkitt's lymphoma cell line) whole cell lysate (10 µg - Lane 2) labeling CIII-Core protein 2 with ab14745 at 5 µg/mL. Secondary antibody is a goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP), 1/3000 dilution.

  • CIV subunit I (ab14705)
    CIV subunit I (ab14705)

    Isolated mitochondria from heart of human (5 µg - Lane 1), cow (1 ug - Lane 2), rat (10 µg - Lane 3), mouse (5 µg - Lane 4) labeling CIV subunit I with ab14705 at 0.5 µg/mL. Secondary antibody is a goat anti-mouse antibody. 

    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.

  • CV alpha subunit (ab14748)
    CV alpha subunit (ab14748)

    Isolated mitochondria from heart of human (10 µg - Lane 1), cow (4 µg - Lane 2), rat (10 µg - Lane 3), mouse (10 µg - Lane 4), HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate (20 µg -Lane 5) labeling CV alpha subunit with ab14748 at 1 µg/mL 

  • CV alpha subunit (ab14748)
    CV alpha subunit (ab14748)

    Human liver tissue lysate, (10 µg - Lane 1) and HepG2 (human liver hepatocellular carcinoma cell line) whole cell  lysate (10 µg - Lane 2) labeling CV alpha subunit with ab14748 at 1 µg/mL. Secondary antibody is a goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP), 1/3000 dilution.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (842)

Publishing research using ab110413? Please let us know so that we can cite the reference in this datasheet.

ab110413 has been referenced in 842 publications.

  • Visavadiya NP  et al. Mitochondrial dynamics and quality control are altered in a hepatic cell culture model of cancer cachexia. Mol Cell Biochem 476:23-34 (2021). PubMed: 32797334
  • Bosoi CR  et al. High-Fat Diet Modulates Hepatic Amyloid ß and Cerebrosterol Metabolism in the Triple Transgenic Mouse Model of Alzheimer's Disease. Hepatol Commun 5:446-460 (2021). PubMed: 33681678
  • Boran T  et al. Extended regorafenib treatment can be linked with mitochondrial damage leading to cardiotoxicity. Toxicol Lett 336:39-49 (2021). PubMed: 33166663
  • Caputo T  et al. Anti-adipogenic signals at the onset of obesity-related inflammation in white adipose tissue. Cell Mol Life Sci 78:227-247 (2021). PubMed: 32157317
  • Currais A  et al. A chemical biology approach to identifying molecular pathways associated with aging. Geroscience 43:353-365 (2021). PubMed: 32705410
View all Publications for this product

Customer reviews and Q&As

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1-10 of 43 Abreviews or Q&A

Oxphos Rodent antibody works on Bovine liver

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Bovine liver 20ug of lysates were loaded on to 4-15% gradient gel.
1:4000 dilution of primary antibody used.
Works great!
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Julie Kim

Verified customer

Submitted Apr 19 2017

Total OXPHOS in whole Gastrocnemius tissue lysate (Mouse)

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Application: WB
Sample species: Mouse
1) Sample species: Mouse
2) Type: Tissue lysate (Gastrocnemius, skeletal muscle)
3) Amount of protein used: 50ug
4) Specification: Skeletal Muscle
5) Reduce/denaturing condition: SDS sample buffer, no boiling. 4-12% gel (Bis/Tris), MOPS
6) Blocking buffer/concentration/time/temperature:Odyssey Blocking Buffer, Antibody dil. in 4%BSA-TBST/4hrs RT
7) Antibody dilution: 1:1000
8) Incubation time/temperature: 4hrs/RT
9) Diluent: 4% BSA-TBST
10) Secondary antibody information:LICOR/Goat-anti-mouse/1:20,000
11) Detection method: LICOR Odyssey
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Alice Pollard

Verified customer

Submitted Apr 08 2015

Total OXPHOS Rodent WB antibody cocktail

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
This kit detected all five components of OXOPHOS complex. The signal was very strong and specific. Highly recommended!
Primary antibody dilution: 1:1000
Secondary antibody (rabbit anti mouse HRP) dilution: 1:3000
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 19 2020

Excellent Antibody!!

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
This antibody is the phenomenon one. You could detect NADH dehydrogenase beta subcomplex subunit 8 of Complex I (NDUFB8), succinate dehydrogenase subunit B of Complex II (SDHB), cytochrome b-c1 complex subunit 2 of Complex III (UQCRC2), Cytochrome c oxidase subunit 1 of Complex IV (MTCO1), and ATP synthase subunit alpha of Complex V (ATP5A) all at once. I used this antibody in 1:1000 dilution -5%BSA. It gives you very nice bands with no background.
Sample: isolated mitochondria from mice brain (control and Alzheimer’s disease (AD)).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Aida Adlimoghaddam

Verified customer

Submitted Jan 15 2020

OXPHOS for Rat endothelial cells

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
Application: Western blot
Sample: Rat Cell lysate - whole cell (Rat microvascular endothelial cells)- triplicates in lanes 2,3,4.
All blue marker in lane 1.
Loading amount: 20 µg
Specification: Rat microvascular endothelial cells
Gel Running Conditions: Reduced Denaturing (12% gel)
Blocking step: Milk as blocking agent for 24 hours · Concentration: 5 % · Temperature: 4°C
Primary antibody conditions:
Incubation time:
1 hour Temperature: 23°C Diluent: TBS+Tween (0.1%)
Secondary antibody
Name: Non-Abcam antibody was used: Goat Anti-Mouse IgG (H+L)-HRP Conjugate
Conjugation: HRP polymer
Dilution:1/5000
Detection method: ECL
Exposure: 1 minute and 0 seconds
Bands: Specific: 50, 40, 30, 25, 18 Kda
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dhanush Haspula

Verified customer

Submitted May 07 2019

OXPHOS Rodent WB antibody cocktail ab110413 in Rabbit samples

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
The molecular weight marker is at 50kDA, 37kDA, 25kDa and 20kDA. In rails 1 to 7 and 9 to 14 are different Rabbit samples (1-3 control rabbit samples) and in rail 8 is the Rat Heart mitochondria WB control. The samples were kept at room temperature (not heated) and a PVDF membrane was used for transfer.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Daniel Jerico

Verified customer

Submitted Apr 12 2019

Total OXPHOS Western Blot C2C12 (mouse)

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Cell line: C2C12 (myoblasts)
Protein loaded 25 ug
Primary antibody dilution: 1:1000
Secondary: 1:10000 (HRP), goat anti-mouse IgG-HRP: sc-2005
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 21 2018

OXPHOS Antibody on Pig Skeletal Muscle

 Average Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Used Pig Skeletal Muscle preps. Loaded 30ug of protein for pig skeletal muscle samples, and 5uL of the Rat Heart Positive Control onto 5-12% gradient gel. Transferred onto nitrocellulose membrane. Blocked with 1XTBST with 5% nonfat dry milk. Antibody was diluted 1:1000 in 1xTBST. Secondary Goat anti Mouse Secondary Antibody (green). After imaging, only picked up 4 bands total in pig muscle (2 very faint).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 02 2018

Question

What do the temperature values in the image represent? The temperature at which the mitochondria was lysed before the electrophoresis? If so, for how long?

Read More
Answer

The temperature shown on the WB image is the degrees at which the lysate was denatured prior to loading on the gel (5 min only). We show this as most customers boil the samples before loading. For these antibodies, boiling will actually be counterproductive as the signal disappears as the temperature is increased.

Read More

Question

Hi, this antibody panel, ab110413, recognizes complex I-V quite sufficiently. I was wondering if there is any difference between the MTCO1 (COX IV subunit I) at 40kDa and another COX IV loading control (ab16056) at 17kDa? Since COX IV is typically used for loading control in the mitochondrial western blot. I use that Ab for other mitochondrial protein analysis. But I am not sure if I can use COX IV in the case of evaluating MTCO1, since they are in some way the same COX IV, right? And why the molecular weight for the proteins recognized by both Abs are different? Thanks! Hailong

Read More
Answer

The antibodies detect different subunits of cytochrome c oxidase, which is also known as Complex IV, and which is abbreviated as CIV on the ab110413 datasheet (Total OXPHOS Rodent WB Antibody Cocktail).

The CIV (Complex IV) antibody in the panel ab110413 detects Compex IV subunit 1, also called COXI and MT-CO1, at ˜40kDa. This antibody is available separately as ab14705. The COX IV mitochondria loading control antibody ab16056 detects Complex IV subunit 4, also known as COXIV, at about 15kDa. Here are links to the UniProt records for the human proteins.

MT-CO1 https://www.uniprot.org/uniprot/P00395
COXIV https://www.uniprot.org/uniprot/P13073

Read More

1-10 of 43 Abreviews or Q&A

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