Product nameMitoTox™ Complex V OXPHOS Activity Assay Kit
See all Complex V kits
Sample typeInhibitor compounds
Assay time4h 00m
Species reactivityReacts with: Rat, Cow, Human
Does not react with: Mouse
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.
Store Phospholipids, Bovine heart mitochondria and Activity Buffer at -80°C. Store all other components at 4°C.
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.
Storage instructionsPlease refer to protocols.
Components 96 tests 12-channel reagent reservoirs 2 units 1X Mito Buffer 1 x 5ml 20X Wash Buffer 1 x 5ml Bovine heart mitochondria 1 x 360µl Complex V Activity Buffer 1 x 24ml Detergent 1 x 100µl Phospholipids 1 x 6ml Pre-coated 96-well microplate 1 unit
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.
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) »