MitoTox Complex I OXPHOS Activity Assay (ab109903) is designed for testing the direct inhibitory effect of compounds on Complex I activity in only 5 hours.
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- The European journal of neuroscience 56:4514-45282022α-Glyceryl-phosphoryl-ethanolamine protects human hippocampal neurons from ageing-induced cellular alterations.Applications:Unspecified applicationReactive species:Unspecified reactive speciesElisa Zappelli,Simona Daniele,Lorenzo Ceccarelli,Matteo Vergassola,Lorella Ragni,Giorgina Mangano,Claudia MartiniPubMed 35902984
- Frontiers in endocrinology 13:8955932022plays an important role in the pathogenesis of metabolic-associated fatty liver disease by regulating hepatic lipid metabolism.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYongqiang Ma,Guangshun Chen,Junfang Yi,Qiang Li,Zhi Tan,Wenling Fan,Xiaohua Luo,Zhiyong He,Zhongzhou Si,Jiequn LiPubMed 35957832
- Cancer discovery 11:2316-23332021NKX3.1 Localization to Mitochondria Suppresses Prostate Cancer Initiation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlexandros Papachristodoulou,Antonio Rodriguez-Calero,Sukanya Panja,Elizabeth Margolskee,Renu K Virk,Teresa A Milner,Luis Pina Martina,Jaime Y Kim,Matteo Di Bernardo,Alanna B Williams,Elvis A Maliza,Joseph M Caputo,Christopher Haas,Vinson Wang,Guarionex Joel De Castro,Sven Wenske,Hanina Hibshoosh,James M McKiernan,Michael M Shen,Mark A Rubin,Antonina Mitrofanova,Aditya Dutta,Cory Abate-ShenPubMed 33893149
- Theranostics 10:12044-120592020ITGB2-mediated metabolic switch in CAFs promotes OSCC proliferation by oxidation of NADH in mitochondrial oxidative phosphorylation system.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiaoxin Zhang et. al.PubMed 33204328
- Food and chemical toxicology : an international jo 137:1111632020FUNDC1 regulates receptor-mediated mitophagy independently of the PINK1/Parkin-dependent pathway in rotenone-treated SH-SY5Y cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSi Yeon Park et. al.PubMed 32001317
- Cell reports 29:1511-1523.e52019Metformin Improves Mitochondrial Respiratory Activity through Activation of AMPK.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYu Wang,Hongying An,Ting Liu,Caolitao Qin,Hiromi Sesaki,Shaodong Guo,Sally Radovick,Mehboob Hussain,Akhil Maheshwari,Fredric E Wondisford,Brian O'Rourke,Ling HePubMed 31693892
- Cancer cell 36:84-99.e82019Mubritinib Targets the Electron Transport Chain Complex I and Reveals the Landscape of OXPHOS Dependency in Acute Myeloid Leukemia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesIrène Baccelli,Yves Gareau,Bernhard Lehnertz,Stéphane Gingras,Jean-François Spinella,Sophie Corneau,Nadine Mayotte,Simon Girard,Mélanie Frechette,Valérie Blouin-Chagnon,Koryne Leveillé,Isabel Boivin,Tara MacRae,Jana Krosl,Clarisse Thiollier,Vincent-Philippe Lavallée,Evgeny Kanshin,Thierry Bertomeu,Jasmin Coulombe-Huntington,Corinne St-Denis,Marie-Eve Bordeleau,Geneviève Boucher,Philippe P Roux,Sébastien Lemieux,Mike Tyers,Pierre Thibault,Josée Hébert,Anne Marinier,Guy SauvageauPubMed 31287994
- Cancer & metabolism 7:22019CHCHD4 confers metabolic vulnerabilities to tumour cells through its control of the mitochondrial respiratory chain.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLuke W Thomas et. al.PubMed 30886710
- Cancer letters 424:127-1352018Pro-apoptotic Bax promotes mesenchymal-epithelial transition by binding to respiratory complex-I and antagonizing the malignant actions of pro-survival Bcl-2 proteins.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEun Mi Kim et. al.PubMed 29596889
- Environmental toxicology 33:1005-10182018NF-κB/p53-activated inflammatory response involves in diquat-induced mitochondrial dysfunction and apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSu Eun Choi et. al.PubMed 29484840
Key facts
- Detection method
- Colorimetric
- Sample types
- Inhibitor compounds
- Assay type
- Quantitative
- Assay time
- 5h
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MitoTox Complex I OXPHOS Activity Assay (ab109903) is designed for testing the direct inhibitory effect of compounds on Complex I activity in only 5 hours.
Key facts
- Detection method
- Colorimetric
- Sample types
- Inhibitor compounds
- Assay type
- Quantitative
- Assay time
- 5h
- 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 I OXPHOS Activity Assay (ab109903) is designed for testing the direct inhibitory effect of compounds on Complex I activity in only 5 hours. Complex I extracted from the provided bovine heart mitochondria (a rich source of Complex I) is immunocaptured by specific antibodies on the plate. Complex I 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 I 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, Ubiquinone and Activity Buffer at -80°C. Store all other components at 4°C.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Complex I also known as NADH dehydrogenase or NADH:ubiquinone oxidoreductase is a large enzyme complex with a molecular mass of approximately 1000 kDa. It is expressed in the inner mitochondrial membrane of eukaryotic cells. As the first enzyme in the mitochondrial respiratory chain Complex I plays a critical mechanical role in cellular respiration. It transfers electrons from NADH to ubiquinone coupled with the translocation of protons across the inner mitochondrial membrane contributing to the generation of a proton gradient used to produce ATP.
- Biological function summary
Complex I acts as an integral component of the mitochondrial respiratory chain which is a series of protein complexes involved in cellular energy production. As part of this complex system Complex I is essential for effective oxidative phosphorylation. Its activity is assessed using protein activity assays including immunocapture or complex activity assays and microplate assays. Complex I activity influences the overall efficiency of ATP production affecting energy-dependent cellular processes.
- Pathways
Complex I functions within the electron transport chain one of the major pathways in cellular respiration. This pathway is vital for ATP synthesis providing the energy currency required by cells. Complex I works closely with other electron transport chain complexes such as Complex II (succinate dehydrogenase complex) and Complex III (cytochrome c reductase) to drive oxidation-reduction reactions and maintain cellular metabolism.
- Associated diseases and disorders
Defects in Complex I are linked with mitochondrial diseases and neurodegenerative disorders such as Leigh syndrome and Parkinson's disease. Mutations in Complex I subunits disrupt normal electron transport and ATP production leading to increased oxidative stress and neuronal cell damage. Other mitochondrial proteins such as cytochrome c oxidase are also implicated in these conditions emphasizing the interconnected nature of mitochondrial dysfunction in disease progression.
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Functional Studies - MitoTox™ Complex I OXPHOS Activity Assay Kit (ab109903)
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.
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