AB109905

MitoTox™ Complex II + III OXPHOS Activity Assay Kit

Name: MitoTox™ Complex II + III OXPHOS Activity Assay Kit (ab109905) | Abcam
Brand: Abcam Limited
SKU: AB109905
Price: 78000 JPY
Availability: InStock

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(42 Publications)

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.

別名を表示する

Complex III subunit 1, Core protein I, Ubiquinol-cytochrome-c reductase complex core protein 1, UQCRC1

2 Images

Functional Studies - MitoTox™ Complex II + III OXPHOS Activity Assay Kit (AB109905)

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PubMed

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 ab109721 (top left), Complex II + III were measured using ab109905 (top right), Complex IV was measured using ab109911 (bottom left) and Complex V was measured using 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

Image courtesy of Yao X et al.PLoS One. 2015; 10(10): e0139416. doi: 10.1371/journal.pone.0139416.

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Supplier Data

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.

Key facts

検出方法

Colorimetric

交差種

Cow, Mammals

アッセイタイプ

Direct

アッセイ時間

30m

アッセイプラットフォーム

Microplate reader

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Reactivity data

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

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製品の詳細

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 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.

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製品内容

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出荷温度及び保存条件

出荷温度

Dry Ice

短期保存温度

Multi

長期保存温度

Multi

保管に関する情報

Please refer to protocols

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補足情報

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

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.

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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製品プロトコール

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ターゲットの情報

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).

See full target information UQCRC1

追加のターゲット

SDHA,SDHB,SDHC,SDHD,MT-CYB,CYC1,Cytochrome b-c1 complex subunit Rieske, mitochondrial,UQCRC2,UQCRH,UQCRB,UQCRQ,UQCR10

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文献 (42)

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

Redox biology 83:103637 PubMed40253748

2025

OXPHOS inhibition overcomes chemoresistance in triple negative breast cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Cemile Uslu,Eda Kapan,Alex Lyakhovich

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

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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

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

Genes & diseases 11:101100 PubMed39281832

2024

Elevated meteorin-like protein from high-intensity interval training improves heart function via AMPK/HDAC4 pathway.

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Yongshun Wang,Jie Yuan,Huadong Liu,Jie Chen,Jieru Zou,Xiaoyi Zeng,Lei Du,Xin Sun,Zhengyuan Xia,Qingshan Geng,Yin Cai,Jingjin Liu

Journal of medical virology 95:e29270 PubMed38047459

2023

SARS-CoV-2 nucleocapsid protein enhances the level of mitochondrial reactive oxygen species.

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Haiyun Yu,Lu Yang,Zhennan Han,Xiaoyu Zhou,Zihan Zhang,Tianli Sun,Fang Zheng,Jingzhi Yang,Fei Guan,Jungang Xie,Chaohong Liu

Scientific reports 13:17176 PubMed37821486

2023

NARFL deficiency caused mitochondrial dysfunction in lung cancer cells by HIF-1α-DNMT1 axis.

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Hongzhou Liu,Xueqin Wu,Tianrong Yang,Chen Wang,Fei Huang,Ying Xu,Jie Peng

The American journal of pathology 193:1528-1547 PubMed37422147

2023

Aberrations in Energetic Metabolism and Stress-Related Pathways Contribute to Pathophysiology in the Neb Conditional Knockout Mouse Model of Nemaline Myopathy.

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Unspecified application

Species

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Rebecca A Slick,Jennifer A Tinklenberg,Jessica Sutton,Liwen Zhang,Hui Meng,Margaret J Beatka,Mark Vanden Avond,Mariah J Prom,Emily Ott,Federica Montanaro,James Heisner,Rafael Toro,Henk Granzier,Aron M Geurts,David F Stowe,R Blake Hill,Michael W Lawlor

Dose-response : a publication of International Hormesis Society 21:15593258231155789 PubMed36798636

2023

Low-Dose Radiation Reduces Doxorubicin-Induced Myocardial Injury Through Mitochondrial Pathways.

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Di Zhao,Xin Jiang,Xinxin Meng,Dandan Liu,Yanwei Du,Lijing Zhao,Hongyu Jiang

Cell metabolism 35:299-315.e8 PubMed36754020

2023

Sodium perturbs mitochondrial respiration and induces dysfunctional Tregs.

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Species

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Beatriz 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 Kleinewietfeld

Chembiochem : a European journal of chemical biology 23:e202200475 PubMed36134475

2022

Identification of Dihydroorotate Dehydrogenase Inhibitors Using the Cell Painting Assay.

Applications

Unspecified application

Species

Unspecified reactive species

Beate 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 Ziegler

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MitoTox™ Complex II + III OXPHOS Activity Assay Kit