
NanoBRET
NanoBRET is used to analyse binding events, signaling pathways and receptor trafficking in live cells and has significantly expanded the range and applications of BRET assays.
The production of energy is an essential cellular function and indicator of cell health. The sources cells use to generate energy can be measured by cell metabolism assays presented here.
The metabolic condition of a cell sheds light on its state of health. The rate of glycolysis is one metabolic factor impacting cellular physiology and pathology. For instance, cancer cells exhibit a higher rate of glycolysis compared to healthy counterparts. Since lactate is produced in the course of glycolysis and subsequently released into the extra-cellular space, it acidifies the surrounding medium. The so-called extracellular acidification is hence used to monitor glycolytic flux.
A second metabolic parameter is the consumption of molecular oxygen (O2) as it reflects oxidative phosphorylation. In cancer cells, this pathway of energy production is decreased. Furthermore, changes in oxygen consumption may indicate mitotoxicity of drugs.
Agilent Technologies provides probes that are sensitive to either pH or O2 and reliably trace both metabolic processes: glycolysis and respiration. Both probes can be used simultaneously, assuring maximal information output from your experiment.
Microplate readers such as the CLARIOstar and VANTAstar are the ideal tools to measure these metabolic assays as they easily detect the probes. and can be equipped with an Atmospheric Control Unit (ACU). This accessory allows working at desired oxygen and carbon dioxide concentrations that resemble physiological conditions. It further enables to work at steady O2 and CO2 concentrations, a requirement for determination of extracellular acidification (ECA) and oxygen consumption rate (OCR).
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NanoBRET is used to analyse binding events, signaling pathways and receptor trafficking in live cells and has significantly expanded the range and applications of BRET assays.
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