MitoXpress® Xtra – Oxygen Consumption Assay [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 MitoXpress® Xtra assay allows measurement of extracellular oxygen consumption rates (OCR) with whole cell populations (both adherent and suspension cells), isolated mitochondria, permeabilised 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 moulds.
In this assay, MitoXpress® Xtra 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 MitoXpress® Xtra nor O2 are consumed), facilitating measurement of time courses and drug treatments.
Luxcel’s flexible plate reader format, allows multiparametric or multiplex combination with Luxcel’s other products, as well as combining with commonly available reagents to measure glycolysis, LDH, JC-1, MMP (Ѱ), ROS, and cellular ATP. For example, MitoXpress® Xtra in combination with Luxcel’s pH-Xtra® – Glycolysis Assay (Cat No. PH-100) allows the simultaneous real-time measurement of mitochondrial respiration and glycolysis and analysis of the metabolic phenotype of cells and the shift (flux) between the two pathways under pathological states.
MitoXpress® Xtra reagent is a chemically stable and inert, bipolymer-based, cell impermeable oxygen sensing fluorophore.
Instrument And Settings
Three fluorescence modalities can be successfully used with the MitoXpress® Xtra – Oxygen Consumption Assay [HS Method], depending on plate reader type and instrument setup, as follow:
1. Basic: Intensity measurement
2. Standard: Time-resolved fluorescence measurement (TR-F), and
3. Advanced: Dual-read Ratiometric TR-F measurement (Lifetime calculation).