SPARC Molecular Analysis makes use of an Agilent Seahorse XFe96 Analyzer to interrogate in real-time the two major energy producing pathways of the cell, mitochondrial respiration and glycolysis. XF technology simultaneously measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), such that cellular functions like oxidative phosphorylation, glycolysis, and fatty acid oxidation, and their responses to substrates, inhibitors, or other compounds can be rapidly determined in a microplate format. The ability to measure the metabolic phenotype of cells and the shift between respiration and glycolysis under pathological conditions will allow you to connect physiological traits of cells with genomic and proteomic data. Researchers have successfully utilized XF technology to gain new insights into diabetes, obesity, cancer, mitochondrial diseases, cardiovascular and neurodegenerative function and more. For more information on XF technology, visit the Seahorse Bioscience website.

Many of our users perform a Glycolysis Stress Test (GST) or Mitochondrial Stress Test (MST) as their first-step, to measure key parameters of mitochondrial function and get a sense of whether further experimentation is warranted. Initially, the optimal cell density needs to be determined to ensure your data is within the optimal range of the XFe96, and FCCP concentration should be optimized for each cell type as well if you are performing a MST (see our Seahorse FAQ to see how we typically do this). Users can also customize their Seahorse assay by, for example, varying cell conditions, injecting different compounds, or determining dose response curves.

 

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