MitoTox™ Complex II+ III OXPHOS Activity Assay Kit (ab109905) is designed for testing the direct inhibitory effect of compounds on Complex III activity in only 30 minutes.
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- Journal of medical virology 95:e292702023SARS-CoV-2 nucleocapsid protein enhances the level of mitochondrial reactive oxygen species.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHaiyun Yu,Lu Yang,Zhennan Han,Xiaoyu Zhou,Zihan Zhang,Tianli Sun,Fang Zheng,Jingzhi Yang,Fei Guan,Jungang Xie,Chaohong LiuPubMed 38047459
- Scientific reports 13:171762023NARFL deficiency caused mitochondrial dysfunction in lung cancer cells by HIF-1α-DNMT1 axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHongzhou Liu,Xueqin Wu,Tianrong Yang,Chen Wang,Fei Huang,Ying Xu,Jie PengPubMed 37821486
- Dose-response : a publication of International Hormesis Society 21:155932582311557892023Low-Dose Radiation Reduces Doxorubicin-Induced Myocardial Injury Through Mitochondrial Pathways.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDi Zhao,Xin Jiang,Xinxin Meng,Dandan Liu,Yanwei Du,Lijing Zhao,Hongyu JiangPubMed 36798636
- Cell metabolism 35:299-315.e82023Sodium perturbs mitochondrial respiration and induces dysfunctional Tregs.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBeatriz F Côrte-Real,Ibrahim Hamad,Rebeca Arroyo Hornero,Sabrina Geisberger,Joris Roels,Lauren Van Zeebroeck,Aleksandra Dyczko,Marike W van Gisbergen,Henry Kurniawan,Allon Wagner,Nir Yosef,Susanne N Y Weiss,Klaus G Schmetterer,Agnes Schröder,Luka Krampert,Stefanie Haase,Hendrik Bartolomaeus,Niels Hellings,Yvan Saeys,Ludwig J Dubois,Dirk Brenner,Stefan Kempa,David A Hafler,Johannes Stegbauer,Ralf A Linker,Jonathan Jantsch,Dominik N Müller,Markus KleinewietfeldPubMed 36754020
- Chembiochem : a European journal of chemical biology 23:e2022004752022Identification of Dihydroorotate Dehydrogenase Inhibitors Using the Cell Painting Assay.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBeate Schölermann,Jana Bonowski,Michael Grigalunas,Annina Burhop,Yusheng Xie,Joseph G F Hoock,Jie Liu,Mark Dow,Adam Nelson,Christine Nowak,Axel Pahl,Sonja Sievers,Slava ZieglerPubMed 36134475
- Frontiers in aging neuroscience 14:8739292022Dihuang-Yinzi Alleviates Cognition Deficits Targeting Energy-Related Metabolism in an Alzheimer Mouse Model as Demonstrated by Integration of Metabolomics and Network Pharmacology.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGuanghui Han,Weizhe Zhen,Yuan Dai,Hongni Yu,Dongyue Li,Tao MaPubMed 35431901
- Frontiers in cell and developmental biology 10:7960612022Evaluation of Parkin in the Regulation of Myocardial Mitochondria-Associated Membranes and Cardiomyopathy During Endotoxemia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMatthew Kim,Azadeh Nikouee,Yuxiao Sun,Qing-Jun Zhang,Zhi-Ping Liu,Qun Sophia ZangPubMed 35265609
- Experimental and therapeutic medicine 23:2362022Long non-coding RNA TUG1 sponges microRNA-9 to protect podocytes from high glucose-induced apoptosis and mitochondrial dysfunction via SIRT1 upregulation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMin Lei,Guibao Ke,Yan Wang,Dan Luo,Yao HuPubMed 35222713
- International journal of molecular sciences 23:2021IL-25 Induced ROS-Mediated M2 Macrophage Polarization via AMPK-Associated Mitophagy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMei-Lan Tsai,Yi-Giien Tsai,Yu-Chih Lin,Ya-Ling Hsu,Yi-Ting Chen,Ming-Kai Tsai,Wei-Ting Liao,Yi-Ching Lin,Chih-Hsing HungPubMed 35008429
- ChemMedChem 17:e2021006422021Movement of Mitochondria with Mutant DNA through Extracellular Vesicles Helps Cancer Cells Acquire Chemoresistance.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEtna Abad,Alex LyakhovichPubMed 34847299
Key facts
- Detection method
- Colorimetric
- Assay type
- Direct
- Reactive species
- Cow, Mammals
- Assay time
- 30m
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Target data
Function
Component of the ubiquinol-cytochrome c oxidoreductase, a multisubunit transmembrane complex that is part of the mitochondrial electron transport chain which drives oxidative phosphorylation. The respiratory chain contains 3 multisubunit complexes succinate dehydrogenase (complex II, CII), ubiquinol-cytochrome c oxidoreductase (cytochrome b-c1 complex, complex III, CIII) and cytochrome c oxidase (complex IV, CIV), that cooperate to transfer electrons derived from NADH and succinate to molecular oxygen, creating an electrochemical gradient over the inner membrane that drives transmembrane transport and the ATP synthase. The cytochrome b-c1 complex catalyzes electron transfer from ubiquinol to cytochrome c, linking this redox reaction to translocation of protons across the mitochondrial inner membrane, with protons being carried across the membrane as hydrogens on the quinol. In the process called Q cycle, 2 protons are consumed from the matrix, 4 protons are released into the intermembrane space and 2 electrons are passed to cytochrome c (By similarity). The 2 core subunits UQCRC1/QCR1 and UQCRC2/QCR2 are homologous to the 2 mitochondrial-processing peptidase (MPP) subunits beta-MPP and alpha-MPP respectively, and they seem to have preserved their MPP processing properties (By similarity). May be involved in the in situ processing of UQCRFS1 into the mature Rieske protein and its mitochondrial targeting sequence (MTS)/subunit 9 when incorporated into complex III (Probable). Seems to play an important role in the maintenance of proper mitochondrial function in nigral dopaminergic neurons (PubMed:33141179).
Additional Targets
SDHA, SDHB, SDHC, SDHD, MT-CYB, CYC1, Cytochrome b-c1 complex subunit Rieske, mitochondrial, UQCRC1, UQCRC2, UQCRH, UQCRB, UQCRQ, UQCR10
Alternative names
Complex III subunit 1, Core protein I, Ubiquinol-cytochrome-c reductase complex core protein 1, UQCRC1
What's included?
Recommended products
MitoTox™ Complex II+ III OXPHOS Activity Assay Kit (ab109905) is designed for testing the direct inhibitory effect of compounds on Complex III activity in only 30 minutes.
Key facts
- Detection method
- Colorimetric
- Assay type
- Direct
- Reactive species
- Cow, Mammals
- Assay time
- 30m
- Assay Platform
- Microplate reader
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- Multi
- Appropriate long-term storage conditions
- Multi
- Storage information
- Please refer to protocols
Notes
MitoTox™ Complex II+ III OXPHOS Activity Assay Kit (ab109905) is designed for testing the direct inhibitory effect of compounds on Complex III activity in only 30 minutes. The assay is performed using whole bovine heart mitochondria, a rich source of OXPHOS complex. Succinate (electron donor of Complex II) and oxidized cytochrome c (electron acceptor of Complex III) are added to the mitochondria to start the electron transfer pathway that takes place during oxidative phosphorylation. The rate of couple Complex II + III reaction is measured by monitoring the conversion of oxidized cytochrome c into reduced form, which can be observed as increase in absorbance at OD 550 nm. The assay requires rotenone and KCN (not supplied): rotenone (Complex I inhibitor) blocks electron transfer to ubiquinone, ensuring that all reduction of cytochrome c happens via Complex II. The addition of KCN (Complex IV inhibitor) ensures that there is no re-oxidation of cytochrome c. The intra-assay and inter-assay variation of this assay are both < 10%.
Inhibitory effects of compounds on Complex III 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 III 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 Bovine Heart Mitochondria and Cytochrome c (oxidized) at -80°C. Store all other components at 4°C. **Related products** Review the , or the full 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Complex III also known as cytochrome c reductase or cytochrome bc1 complex plays an important role in the mitochondrial electron transport chain. This multiprotein complex weighing approximately 250 kDa facilitates electron transfer from ubiquinol to cytochrome c. Complex III resides in the inner mitochondrial membrane of eukaryotic cells and is important for cellular energy production. Structurally it consists of 11 subunits in mammals with cytochrome b cytochrome c1 and Rieske iron-sulfur protein being the core components.
- Biological function summary
Complex III transfers electrons and contributes to proton translocation across the mitochondrial membrane. This process generates the proton gradient needed for ATP synthesis via oxidative phosphorylation often referred to as OXPHOS. As a core component of the mitochondrial respiratory chain Complex III interacts closely with other complexes mainly Complexes I and IV to maintain efficient energy conversion.
- Pathways
Complex III is integral to the oxidative phosphorylation pathway and the respiratory electron transport chain. It ensures efficient electron transport working closely with Complexes I II and IV. Cytochrome c acts as an electron carrier between Complex III and Complex IV in this pathway. This coordinated interaction is essential for ATP generation meeting cellular energy demands.
- Associated diseases and disorders
Complex III is linked with mitochondrial disorders and neurodegenerative diseases. Deficiencies or mutations in its components may lead to disorders such as mitochondrial encephalopathy or Leigh syndrome which are associated with impaired energy metabolism. The complex's dysfunction also affects proteins connected to these diseases like ATP synthase due to disrupted energy production. Additionally its role in oxidative stress has implications for conditions like mitotox-related mitochondrial damage.
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2 product images
Image courtesy of Yao X et al.PLoS One. 2015; 10(10): e0139416. doi: 10.1371/journal.pone.0139416.Functional Studies - MitoTox™ Complex II + III OXPHOS Activity Assay Kit (ab109905)
Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis.
Mitchondiral fractions from the heart tissue of Rats infected by S. pneumoniae, or given PBS sham control, were subjected to measurements of complex I-V activities. Complex I was measured with Complex I Enzyme Activity Microplate Assay Kit (Colorimetric) ab109721 (top left), Complex II + III were measured using ab109905 (top right), Complex IV was measured using Complex IV Rodent Enzyme Activity Microplate Assay Kit ab109911 (bottom left) and Complex V was measured using ATP synthase Activity Assay Kit (Colorimetric) ab109714 (bottom right).
Freshly isolated mitochondrial pellets were resuspened in PBS supplemented with 10% detergent provided in the kits. Protein concentrations of these mitochondrial lysates were estimated and 25 μg (for complex I, IV and V) or 100 μg (for complex II+III) mitochondrial protein was used per reaction. Enzyme activities were measured spectrophotometricly in triplicate and expressed as changes of absorbance per minute per mg protein
Functional Studies - MitoTox™ Complex II + III OXPHOS Activity Assay Kit (ab109905)
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.
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