MitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays) (ab110419)

Overview

  • Product nameMitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays)
  • Detection methodColorimetric
  • Sample type
    Inhibitor compounds
  • Assay typeQuantitative
  • Species reactivity
    Reacts with: Cow, Human
  • Product overview

    MitoTox™ Complete OXPHOS Activity Assay Kit (5 assays) (ab110419) is designed for testing the direct inhibitory effect of compounds on the 5 OXPHOS complexes. Inhibitory effects of compounds on OXPHOS Complexes activity can be tested in two different ways: 1. Screening format, where up to 23 compounds can be tested at a single concentration in triplicate; 2. Dose response (IC50) format, where two compounds known to affect specific Complex activity can be tested at 11 different data points in triplicate.


    Complexes I, II, IV and V activity are each immunocaptured from bovine heart mitochondria by specific antibodies on 96-well microplates, and their activities measured by simple spectrophotometric assays. Complex III activity is measured by an in-solution colorimetric assay.


    Testing for mitochondrial function has become a key aspect of drug discovery. Mitochondria can be affected by drug treatment, resulting into cardio- and hepatotoxic side effects that can lead to drug withdrawal from the market. Therefore, there is increasing emphasis on testing the impact on mitochondria early on in the drug development process to reduce failure rates during preclinical and clinical phases. 

  • Notes

    This kit consists of the following assays:

    MitoTox™ Complex I OXPHOS Activity Assay Kit (ab109903)

    MitoTox™ Complex II OXPHOS Activity Assay Kit (ab109904)

    MitoTox™ Complex II + III OXPHOS Activity Assay Kit (ab109905)

    MitoTox™ Complex IV OXPHOS Activity Assay Kit (ab109906)

    MitoTox™ Complex I OXPHOS Activity Assay Kit (ab109907)

  • Tested applicationsSuitable for: Functional Studiesmore details
  • PlatformMicroplate

Properties

  • Storage instructionsPlease refer to protocols.
  • Components 5 x 96 tests
    10X Detergent 1 x 1ml
    12-channel reagent reservoirs 10 units
    1X Blocking Buffer (Solution 1) 1 x 70ml
    1X Complex III Mito Buffer 1 x 1ml
    1X Complex IV Mito Buffer (Solution 2) 1 x 110ml
    1X Mito Buffer 2 x 5ml
    1X Succinate Solution 1 x 12ml
    20 x Buffer 1 x 5ml
    20X Wash Buffer 2 x 5ml
    96-well microplate 1 unit
    Bovine heart mitochondria 4 x 360µl
    Bovine Heart Mitochondria 1 x 90µl
    Bovine heart mitochondria (5 mg/mL) 1 x 300µl
    Complex I Activity Buffer 1 x 24ml
    Complex II Activity Buffer 1 x 24ml
    Complex V Activity Buffer 1 x 24ml
    Cytochrome c (oxidized) 1 x 550µl
    DCIP 1 x 250µl
    Detergent 1 x 20µl
    Detergent 2 x 100µl
    Detergent (ab109906) 1 x 20µl
    Fat free powdered milk (Blocking powder) 1 x 1.8g
    Phospholipid 2 x 6ml
    Pre-coated 96-well microplate 1 unit
    Pre-coated 96-well microplate 1 unit
    Pre-coated 96-well microplate 1 unit
    Pre-coated 96-well microplate 1 unit
    Reagent C (Cytochrome c) 2 x 1ml
    Single-channel reagent reservoirs 2 units
    Succinate 1 x 500µl
    Ubiquinone 1 1 x 60µl
    Ubiquinone 2 1 x 60µl
  • Research areas

    Applications

    Our Abpromise guarantee covers the use of ab110419 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
    Functional Studies Use at an assay dependent dilution.

    MitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays) images

    • Dose response curve for aurovertin, a non-competitive inhibitor of Complex V.

    • Typical dose response curve for oligomycin. Assay was performed following the Dose Response Assay Procedure using oligomycin, a well known Complex V inhibitor. Oligomycin was prepared in DMSO to generate a 10 mM stock. Starting with a 50 µM final concentration in well, 1:10 serial dilutions of oligomycin were generated.

    • Typical dose response curve for KCN. Assay was performed following the Dose Response Assay Procedure using KCN, a well known Complex IV inhibitor. KCN was prepared in 0.1 M NaOH to generate a 100 mM stock. Starting with a 100 µM final concentration in well, 1:10 serial dilutions of KCN were generated.

    • Typical dose response curve for antimycin A. Assay was performed following the Dose Response Assay Procedure using antimycin A, a well known Complex III inhibitor. Antimycin A was prepared in DMSO to generate a 10 mM stock. Starting with a 150 µM final concentration in well, 1:4 serial dilutions of antimycin A were generated.

    • Typical dose response curve for TTFA (2-thenoyltrifluoracetone). Assay was performed following the Dose Response Assay Procedure using TTFA, a well known Complex II inhibitor. TTFA was prepared in DMSO to generate a 100 mM stock. Starting with a 500 µM final concentration in well, 1:2 serial dilutions of TTFA were generated.

    • Typical dose response curve for rotenone. Assay was performed following the Dose Response Assay Procedure using rotenone, a well known Complex I inhibitor. Rotenone was prepared in DMSO to generate a 10 mM stock. Starting with a 50 µM final concentration in well, 1:10 serial dilutions of rotenone were generated.

    Protocols

    References for MitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays) (ab110419)

    This product has been referenced in:
    • Lai K  et al. Integrated Compound Profiling Screens Identify the Mitochondrial Electron Transport Chain as the Molecular Target of the Natural Products Manassantin, Sesquicillin, and Arctigenin. ACS Chem Biol N/A:N/A (2012). Read more (PubMed: 23138533) »

    See 1 Publication for this product

    Product Wall

    On our datasheet we state that this kit can be used with human and cow.

    Therefore yes I can confirm that you can use it on your human samples. What I think may be confusing is that we supply bovine heart mitochondria rather than human.
    Read More


    It is crucial that this type of data is analyzed based on the actual kinetic trace (via softmax software). When a reduction trace is superimposed you will notice that the kinetic change in signal is not constant for the entire assay. The speed of ...

    Read More

    The mitotox kits will work with mitochondria from any human tissue. Therefore it will work with mitochondria from human PBMCs.


    To isolate mitochondria from human PBMCs I suggest the customer to use ab110170 (Mitochondria Isolation Kit f...

    Read More

    The kits that are the best suited for your experiment are listed below. I am having our datasheets team update the species reactivity.
    ab109903, Complex I; reactivity = Human, mouse, rat, bovine
    http://www.abcam.com/mitotox%E2%84%A2-complex-i...

    Read More

    Thank you for contacting Abcam.

    MitoSciences is part of the Abcam family now, and I have directly contacted them regarding your inquiry. They responded that the 384 well plate format has not been validated or produced. Since Abcam and MitoScie...

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    Thank you for contacting Abcam.

    I contacted the supplier, and they responded that ab110419 has not been validated in 384 well plate format.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need an...

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    Thank you for contacting Abcam.

    We only make custom products for bulk orders of 10 or more products. If you would like to inquire about placing a bulk order for ab110419 in 384-well format please reply to this email, and we will see if the su...

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    Thank you for contacting us.
    The assays measure the direct effects of drugs on the 5 OXPHOS complexes and can be used on human samples.

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

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    I now have further information to share with you.

    The antibodies used in the MitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays), ab110419,to coat the plates are cross-reactive againstthe human proteins. The kit can therefore be used...

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    Thank you for contacting us and your interest in our products.

    The MitoTox™ Complete OXPHOS Activity Assay Kit (5 Assays), ab110419, provides bovine heart mitochondria to allow for complexes I, II, IV and V to be immunecaptured and thei...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"