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AB109903

MitoTox™ Complex I OXPHOS Activity Assay Kit

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(20 Publications)

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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Functional Studies - MitoTox™ Complex I OXPHOS Activity Assay Kit (AB109903)
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Supplier Data

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.

Key facts

Detection method

Colorimetric

Sample types

Inhibitor compounds

Results type

Quantitative

Assay time

5h

Assay Platform

Microplate reader

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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.

What's included?

{ "values": { "96Test": { "sellingSize": "96 Test", "publicAssetCode":"ab109903-96Test", "assetComponentDetails": [ { "size":"1 x 360 µL", "name":"Bovine heart mitochondria", "number":"AB109903-CMP07", "productcode":"" }, { "size":"1 x 6 mL", "name":"Phospholipids", "number":"AB109903-CMP09", "productcode":"" }, { "size":"1 x 24 mL", "name":"Complex I Activity Buffer", "number":"AB109903-CMP08", "productcode":"" }, { "size":"1 x 5 mL", "name":"20X Wash Buffer", "number":"AB109903-CMP05", "productcode":"" }, { "size":"2 x 1 Unit", "name":"12-channel reagent reservoirs", "number":"AB109903-CMP03", "productcode":"" }, { "size":"1 x 100 µL", "name":"Detergent", "number":"AB109903-CMP02", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Pre-coated 96-well microplate", "number":"AB109903-CMP06", "productcode":"" }, { "size":"1 x 60 µL", "name":"Ubiquinone 1", "number":"AB109903-CMP01", "productcode":"" }, { "size":"1 x 5 mL", "name":"1X Mito Buffer", "number":"AB109903-CMP04", "productcode":"" } ] } } }

Properties and storage information

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
Multi
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

Product protocols

Target data

Publications (20)

Recent publications for all applications. Explore the full list and refine your search

Cell death & disease 16:489 PubMed40610445

2025

The dual mechanism of mA demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaoya Sun,Qinmei Tan,Yue Yang,Jia Wei,Xiaodie Zhou,Shanshan Gao,Fei Yang

Frontiers in physiology 16:1554222 PubMed40313872

2025

Effects of high-intensity interval training and moderate-intensity continuous training on mitochondrial dynamics in human skeletal muscle.

Applications

Unspecified application

Species

Unspecified reactive species

Yuqing Li,Wanjun Zhao,Qi Yang

Cell death discovery 11:15 PubMed39828731

2025

CRAT downregulation promotes ovarian cancer progression by facilitating mitochondrial metabolism through decreasing the acetylation of PGC-1α.

Applications

Unspecified application

Species

Unspecified reactive species

Zhen Zhang,Shuhua Zhao,Xiaohui Lv,Yan Gao,Qian Guo,Yanjie Ren,Yuanyuan He,Yihua Jin,Hong Yang,Shujuan Liu,Xiaohong Zhang

Science advances 10:eadq0101 PubMed39453997

2024

macrophage migration inhibitory factor shows anti- potential via AZIN1/STAT1 interaction.

Applications

Unspecified application

Species

Unspecified reactive species

Chanjin Yoon,Hyo Keun Kim,Yu Seong Ham,Woo Jin Gil,Seok-Jun Mun,Euni Cho,Jae-Min Yuk,Chul-Su Yang

CytoJournal 21:22 PubMed38989294

2024

Mitochondrial protein isoleucyl-tRNA synthetase 2 in tumor cells as a potential therapeutic target for cervical cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaojiao Meng,Bo Gao,Ning Li

Clinical and experimental pharmacology & physiology 51:e13860 PubMed38584327

2024

NPAS2, transcriptionally activated by ARRB1, promotes the malignant behaviours of lung adenocarcinoma cells and regulates the reprogramming of glucose metabolism.

Applications

Unspecified application

Species

Unspecified reactive species

Shenglan Wang,Chunhong Huang,Yanbin Zheng,Xinjie Wu,Yutong Zhong

The European journal of neuroscience 56:4514-4528 PubMed35902984

2022

α-Glyceryl-phosphoryl-ethanolamine protects human hippocampal neurons from ageing-induced cellular alterations.

Applications

Unspecified application

Species

Unspecified reactive species

Elisa Zappelli,Simona Daniele,Lorenzo Ceccarelli,Matteo Vergassola,Lorella Ragni,Giorgina Mangano,Claudia Martini

Frontiers in endocrinology 13:895593 PubMed35957832

2022

plays an important role in the pathogenesis of metabolic-associated fatty liver disease by regulating hepatic lipid metabolism.

Applications

Unspecified application

Species

Unspecified reactive species

Yongqiang Ma,Guangshun Chen,Junfang Yi,Qiang Li,Zhi Tan,Wenling Fan,Xiaohua Luo,Zhiyong He,Zhongzhou Si,Jiequn Li

Cancer discovery 11:2316-2333 PubMed33893149

2021

NKX3.1 Localization to Mitochondria Suppresses Prostate Cancer Initiation.

Applications

Unspecified application

Species

Unspecified reactive species

Alexandros 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-Shen

Theranostics 10:12044-12059 PubMed33204328

2020

ITGB2-mediated metabolic switch in CAFs promotes OSCC proliferation by oxidation of NADH in mitochondrial oxidative phosphorylation system.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaoxin Zhang,Yingchun Dong,Mengxiang Zhao,Liang Ding,Xihu Yang,Yue Jing,Yuxian Song,Sheng Chen,Qingang Hu,Yanhong Ni
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