Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sample type: Adherent cells, Purified mitochondria, Suspension cells, Tissue
Product nameExtracellular Oxygen Consumption Assay
See all Extracellular Oxygen Consumption kits
Sample typeTissue, Adherent cells, Suspension cells, Purified mitochondria
Assay time1h 30m
Extracellular Oxygen Consumption Assay Kit ab197243 is a mix-and-read, 96-well fluorescence plate reader assay for the real-time kinetic analysis of extracellular oxygen consumption rates (OCR). The oxygen consumption rate is a measure of the cellular respiration rate, and of mitochondrial function.
The assay is optimized for isolated mitochondria and cell cultures, and can be used with tissues, enzyme preparations, and small organisms.
The fluorescent dye used in this assay kit is quenched by oxygen. The dye excites at 360-380 nm (max 380) and emits at 630-680 nm (max 650). It is also available separately as ab197242.
In the assay, an oil layer is added on top of the assay medium to limit diffusion of oxygen into the assay medium. As mitochondrial respiration depletes the oxygen within the assay medium, quenching of the fluorescent dye is reduced, and the fluorescence signal increases proportionately.
The reaction is non-destructive and fully reversible (the oxygen sensitive dye is not consumed) enabling assay time courses and drug treatments.
Learn more about the full range of assays to measure glycolysis, oxygen consumption, fatty acid oxidation and metabolic flux in live cells.
Or review the full metabolism assay guide for other assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 96 tests 4 x 96 tests Extracellular O2 Consumption Reagent 1 vial 4 vials High Sensitivity Oil 1 x 15ml 4 x 15ml
- Extracellular Oxygen Consumption Reagent (ab197242)
- Glycolysis Assay [Extracellular acidification] (ab197244)
- Intracellular Oxygen Concentration Assay (ab197245)
- Mitochondrial Stress Test Complete Assay Kit (ab232857)
- Mitochondrial Stress Test Companion Assay (ab243390)
- Mineral Oil High sensitivity (ab243855)
Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 µM] and FCCP [2.5 µM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).
Typical Lifetime profile of Extracellular O2 Consumption Assay for adherent cells, treated with different ETC compounds, including Antimycin A (recommended as a Negative Control). The effect of Glucose Oxidase as a positive Signal Control is illustrated schematically.
Excitation and emission spectra of Extracellular O2 Consumption Reagent. Left panel shows normalized excitation (Ex = 360-400nm; Peak 380nm). Right panel shows emission (Em = 630 - 680nm; Peak 650nm) in oxygenated and deoxygenated conditions.
ab197243 has been referenced in 73 publications.
- Rebollo-Hernanz M et al. Phytochemicals from the Cocoa Shell Modulate Mitochondrial Function, Lipid and Glucose Metabolism in Hepatocytes via Activation of FGF21/ERK, AKT, and mTOR Pathways. Antioxidants (Basel) 11:N/A (2022). PubMed: 35052640
- Guo Q et al. Mitochondrial pyruvate carrier blockade results in decreased osteoclastogenesis and bone resorption via regulating mitochondrial energy production. J Biol Chem N/A:101775 (2022). PubMed: 35257748
- Rebollo-Hernanz M et al. Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro. Front Nutr 9:866233 (2022). PubMed: 35392289
- Kuang Y et al. NADH dehydrogenase subunit 1/4/5 promotes survival of acute myeloid leukemia by mediating specific oxidative phosphorylation. Mol Med Rep 25:N/A (2022). PubMed: 35425997
- Fang Y et al. LAMC1-mediated preadipocytes differentiation promoted peritoneum pre-metastatic niche formation and gastric cancer metastasis. Int J Biol Sci 18:3082-3101 (2022). PubMed: 35541892