Overview

  • Product name
    MitoTox™ Complex V OXPHOS Activity Assay Kit
    See all Complex V kits
  • Detection method
    Colorimetric
  • Sample type
    Inhibitor compounds
  • Assay type
    Direct
  • Assay time
    4h 00m
  • Species reactivity
    Reacts with: Rat, Cow, Human
    Does not react with: Mouse
  • Product overview

    MitoTox™ Complex V OXPHOS Activity Assay Kit (ab109907) is designed for testing the direct inhibitory effect of compounds on Complex V activity in only 4 hours. Complex V extracted from the provided bovine heart mitochondria (a rich source of Complex V) is immunocaptured by specific antibodies on the plate. Complex V activity can be observed as decrease in absorbance at OD 340 nm. The intra-assay and inter-assay variation of this assay are both < 10%.


    Inhibitory effects of compounds on Complex I 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 Complex V activity can be tested at 11 different data points in triplicate.


    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

    Store Phospholipids, Bovine heart mitochondria and Activity Buffer at -80°C. Store all other components at 4°C.

  • Platform
    Microplate

Properties

Images

  • MitoTox™ Complex V OXPHOS Activity Assay (ab109907)
    MitoTox™ Complex V OXPHOS Activity Assay (ab109907)

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

  • MitoTox™ Complex V OXPHOS Activity Assay (ab109907)
    MitoTox™ Complex V OXPHOS Activity Assay (ab109907)

    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.

Protocols

References

This product has been referenced in:
  • Goldberg J  et al. The mitochondrial ATP synthase is a shared drug target for aging and dementia. Aging Cell 17:N/A (2018). Mouse . Read more (PubMed: 29316249) »
  • Kim YR  et al. Toxoplasma gondii GRA8 induces ATP5A1-SIRT3-mediated mitochondrial metabolic resuscitation: a potential therapy for sepsis. Exp Mol Med 50:e464 (2018). Read more (PubMed: 29869623) »

See all 9 Publications for this product

Publishing research using ab109907? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Submit a review Submit a question

7 reviews or Q&A

10
Votes: Highest First

Hello
I am interested in your product ab109907. However it seems to give reading of ATPase activity through measuring reverse mode. Would you have any suggestions as to means by which we could measure forward mode of the ATPase specifically with a...

Read More

Thank you for contacting us.

Unfortunately none of our products measure the forward reaction. As far as I know there are no companies selling kits measuring the forward reaction of the ATP synthase I am afraid.

I am sorry I could no...

Read More

I am looking for Complex I-IV activity kits to study in vitro activity inhibitors after incubating mouse, rat, and human cells with the inhibitors prior to mitochondrial extraction. I am worried that either the kit will not detect the inhibited portion...

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
https://www.abcam.com/mitotox%E2%84%A2-complex-...

Read More



Inquiry: Hi, How is the bovine heart mitochondria in this kit prepared? What buffer is it in when it is shipped to us? Thanks.

Thank you for contacting us.

The bovine mitochondria is in Tris, sucrose, EDTA buffer pH 7.8


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

Product code: 109714 Inquiry: Hi Abcam, Do you know if this kit will immunocapture the ATP synthase enzyme from C. elegans? Thanks!

Product code: 109715: Inquiry: Hi Abcam, Do you know if this kit will capture ATP synthase from C. elegans? T...

Read More

Product code: 109714 Inquiry: Hi Abcam, Do you know if this kit will immunocapture the ATP synthase enzyme from C. elegans? Unlikely unfortunately – this kit was tested positive with human, rat and cow. Mouse was negative. I think it is unl...

Read More

Will this kit work with mouse samples?

Unfortunately not. It comes with a sample (bovine) which cannot be substituted for mouse.

I need some help doing the analysis using this kit. For example I have a high absorbance reading of 0.37 and low abs reading of 0.25. Putting this into the equation in the data analysis section of the protocol shows 0.12/18 minutes. I'm not sure how to...

Read More

The rate quoted in the booklet is mOD/min. I think this customer is quoting OD. Therefore 0.12/18 is 0.0067 OD/min = 6.7 mOD/min

Changing the window analyzed to identify the steepest straight line slope is necessary and then to compare all sa...

Read More

Hi Abcam, I ordered the MitoTox™ Complex V OXPHOS Activity Microplate Assay (ab109907) before. It works great. However, I needed to include a "negative control" in my work specifically, I wanted to have a compound that does not effect t...

Read More

Thank you for your enquiry.

The negative control chosen for ab109907 MitoTox™ Complex V OXPHOS Activity Microplate Assaywould depend on whatyou want to accomplish and obtain from the experiment.

Some people test the specific...

Read More

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

Sign up