Key features and details
- Assay type: Direct
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr
- Sample type: Inhibitor compounds
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.
ab109907 has been referenced in 15 publications.
- Kim JS et al. Toxoplasma gondii GRA8-derived peptide immunotherapy improves tumor targeting of colorectal cancer. Oncotarget 11:62-73 (2020). PubMed: 32002124
- Baccelli I et al. Mubritinib Targets the Electron Transport Chain Complex I and Reveals the Landscape of OXPHOS Dependency in Acute Myeloid Leukemia. Cancer Cell 36:84-99.e8 (2019). PubMed: 31287994
- Li J et al. Melatonin attenuates renal fibrosis in diabetic mice by activating the AMPK/PGC1a signaling pathway and rescuing mitochondrial function. Mol Med Rep 19:1318-1330 (2019). PubMed: 30535482
- Dehghan E et al. Hydralazine targets cAMP-dependent protein kinase leading to sirtuin1/5 activation and lifespan extension in C. elegans. Nat Commun 10:4905 (2019). PubMed: 31659167
- Wang Y et al. Metformin Improves Mitochondrial Respiratory Activity through Activation of AMPK. Cell Rep 29:1511-1523.e5 (2019). PubMed: 31693892