Cell Metabolism: MitoXpress and pH-Xtra Assays

Agilent’s MitoXpress and pH-Xtra reagent kits enable simple mix and measure assays that utilize soluble, fluorescent nano sensor probes to measure oxygen consumption rates (OCR) and extracellular acidification rates (ECAR). These sensors were designed to provide maximum ease-of-use, higher throughput, and multiplexed data markers for kinetic metabolism assays.

Combined with BioTek’s range of sensitive and versatile microplate readers, this system provides a complete solution optimized for measuring cell metabolism in live cells. Custom designed filter cubes and advanced detection modes reduce background and increase signal dynamic range. CO2/O2 control, incubation, dual reagent dispensers, and direct bottom illumination ensure optimal performance for live-cell assays.  

Our application note titled “Optimization of a Multi-Mode Detection Model for Measuring Real-Time Cellular Respiration and Mitochondrial Function using Fluorescent Biosensors” provides detailed methods, including pre-configured protocols and User Tutorials, for optimizing MitoXpress and pH-Xtra assays across a range of BioTek instruments and conditions.

Read the full application note to discover how combining MitoXpress and pH-Xtra reagents with BioTek plate readers provide sensitive detection methods, including advanced lifetime (µs) time-resolved fluorescence (TRF), for getting the most meaningful data out of your cell metabolism assays.

 

Optimization of a Multi-Mode Detection Model for Measuring Real-Time Cellular Respiration and Mitochondrial Function using Fluorescent Biosensors

Figure (A.) Kinetic lifetime detection of extracellular oxygen consumption of HepG2 cells treated with uncoupling and inhibitor compounds at an interval of 2 min 19 secs. FCCP is a mitochondrial oxidative phosphorylation uncoupler that drives cellular respiration to a maximal rate increasing the depletion of oxygen. AntiMycin blocks cellular respiration via inhibition of complex III of the mitochondrial electron transport chain (ETC) resulting in low to no reduction of oxygen. Slope on lifetime was calculated from the linear portion (10-80 mins) of the 2-hour kinetic run for the OCR calculation. (B.) OCR profiling of the uncoupling and inhibition agents in comparison to untreated cells.

 

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