Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Purified mitochondria, Suspension cells, Tissue
Product nameMitochondrial Stress Test Complete Assay Kit
See all Extracellular Oxygen Consumption kits
Sample typeTissue, Adherent cells, Suspension cells, Purified mitochondria
Assay durationMultiple steps standard assay
Mitochondrial Stress Test Complete Assay Kit (ab232857) (HS Method) is a highly flexible 96- or 384-well fluorescence plate reader-based approach, for the direct, real-time analysis of cellular respiration and mitochondrial function.
The easy-to-use assay allows measurement of extracellular oxygen consumption rates (OCR) with whole cell populations (both adherent and suspension cells), isolated mitochondria, permeabilized cells and a wide range of 3D cultures including: tissues, small organisms, spheroids, scaffolds and matrixes. The assay is also suitable for measurement of isolated enzymes, bacteria, yeasts and molds.
The Extracellular O2 probe is chemically stable and inert, water-soluble and cell impermeable, making it the ideal and scalable mix-and-measure reagent for use in a wide range of cell culture conditions - all measured using a fluorescence plate-reader. In this assay, Extracellular O2 probe is quenched by O2, through molecular collision, and thus the amount of fluorescence signal is inversely proportional to the amount of extracellular O2 in the sample. Rates of oxygen consumption are calculated from the changes in fluorescence signal over time.
The reaction is non-destructive and fully reversible (neither Extracellular O2 probe nor O2 are consumed), facilitating measurement of time courses and drug treatments.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests Antimycin A 1 x 3µg Extracellular O2 probe 1 vial FCCP 1 x 4µg Glucose Oxidase 1 x 112.5µg HS Mineral Oil 1 x 15ml Oligomycin 1 x 10µg
Using this plate map, up to 7 different conditions can be tested simultaneously in triplicate across all treatment conditions.
Slopes (m) calculated from the linear portion of these signal profiles are reflective of the OCR under the conditions imposed by the kit components and are used to determine the key characteristics of aerobic respiration.
a) The OCR (m) from the linear portion of the signal profile from each Stress Test condition reflect Basal Respiration, Non ATP-Coupled Oxygen Consumption, Maximal Respiration and Non Respiratory Oxygen Consumption. ATP-Coupled Oxygen Consumption and Spare Respiratory Capacity are calculated from these values.
b) The contribution of each of these discrete metabolic processes to the Maximal Respiration can be conveniently visualized producing a detailed picture of aerobic metabolism.
In low glucose medium, Spare Respiratory Capacity is low, but significantly increases in the presence of high glucose concentrations or 150 µM Oleate, due to the provision of reducing equivalents to fuel ETC activity.
ab232857 has not yet been referenced specifically in any publications.