ジャーナル
/
生化学
/
Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
JoVE Journal
生化学
Author Produced
このコンテンツを視聴するには、JoVE 購読が必要です。  サインイン又は無料トライアルを申し込む。
JoVE Journal 生化学
Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

DOI:
10.3791/56975-v

•

08:57 min

•

February 24, 2018

•

, , ,
1Department of Biochemistry,Memorial University of Newfoundland

章

  • 00:00標題
  • 00:17Introduction
  • 01:50Setting Up the Assay
  • 05:44Results: Simultaneous tracking of changes in NADH and superoxide/hydrogen peroxide fluorescence
  • 07:32Conclusion

概要

自動翻訳

自動翻訳

Mitochondria contain several flavin-dependent enzymes that can produce reactive oxygen species (ROS). Monitoring ROS release from individual sites in mitochondria is challenging due to unwanted side reactions. We present an easy, inexpensive method for direct assessment of native rates for ROS release using purified flavoenzymes and microplate fluorometry.

タグ

Mitochondrial DehydrogenasesSuperoxide/hydrogen PeroxideNADH ProductionFlavin-containing EnzymesPyruvate DehydrogenaseAlpha-ketoglutarate DehydrogenaseMicroplate FluorometryReactive Oxygen SpeciesAntioxidantsInhibitorsSimultaneous Measurement

Article

Embed

プレイリスト追加

Usage Statistics

関連ビデオ

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Laboratory Scale Production and Purification of a Therapeutic Antibody

Analyses of Mitochondrial Calcium Influx in Isolated Mitochondria and Cultured Cells

Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue

Measuring Liver Mitochondrial Oxygen Consumption and Proton Leak Kinetics to Estimate Mitochondrial Respiration in Holstein Dairy Cattle

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy

Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

Assessing Mitochondrial Function in Sciatic Nerve by High-Resolution Respirometry

Read Article