Bioenergetics Phenotyping Core
University of Mississippi

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The Seahorse analyzer simultaneously measures oxygen and pH in real-time to determine mitochondrial respiration (oxidative phosphorylation; OxPhos) via the oxygen consumption rate (OCR), and extracellular acidification rate (ECAR). Using these measurements, the total Proton Efflux Rate (PER) can be calculated as an accurate measurement of glycolysis rates. These aspects of mitochondrial function and glycolysis are critical for cellular homeostasis and are perturbed in many disorders including aging, cancer, cardiovascular dysfunction, diabetes, obesity, metabolic disorders, neurodegeneration, and viral challenge. Seahorse Analyzers are compatible with a variety of translational disease models, including primary cells and cell lines (adherent or suspension cells), isolated mitochondria, islets, 3-D spheroids, zebrafish embryos, C. elegans, D. melanogaster, yeast, tissue explants, and other biologicals.

Seahorse Analyzer

Seahorse XFe96 Reagents and Materials

Users operate the Seahorse analyzer with their own materials. Assay materials can be acquired from Agilent Technologies (Agilent Seahorse Analyzer) or Fisher Scientific (Fishersci). At these sites, users may purchase all of the plastic ware, reagents, and assay kits needed.

Plastic ware for the XFe96 (Plastic ware brochure)

  • Seahorse XFe96 FluxPak (Agilent #102416-100)
    • Contents: 18 XFe96 Sensor Cartridges, 20 XF96 V3 PS Tissue Culture Microplates
  • Seahorse XFe96 FluxPak mini (Agilent #102601-100)
    • Contents: 6 XFe96 Sensor Cartridges, 10 XF96 V3 PS Tissue Culture Microplates
  • Seahorse XFe96 Spheroid FluxPak (Agilent #102905-100)
    • Contents: 6 XFe96 Sensor Cartridges, 6 XFe96 Spheroid Culture Micoplates
  • Seahorse XF96 V3 PS Culture Microplates (Agilent #101085-004)
    • Contents: 10 Polystyrene plates
  • Seahorse XFe96 Spheroid Plate (Agilent #102959-100)
    • Contents: 1 Polystyrene plate. 1mm (divot diameter)
  • Seahorse XFe96 Spheroid Microplates (Agilent #102978-100)
    • Contents: 6 Polystyrene plates. 1mm (divot diameter)

Reagents/Media for the XFe96

Assay kits for the XFe96 (Agilent Seahorse Consumables)

  • Real-Time ATP Rate Assay Kit (Agilent #103592-100)
    • ATP rates from OxPhos and glycolosis (via oligomycin, rotenone/antimycin A)
  • Mitochondrial Stress Test Kit (Agilent #103015-100)
    • ATP-linked respiration, proton leak, maximal respiration, spare respiratory capacity, and non-mitochondrial respiration (via oligomycin, FCCP, and rotenone/antimycin A)
  • Glycolytic Rate Assay Kit (Agilent #103344-100)
    • precise glycolysis and rapid metabolic switches not discernible in endpoint lactate assays (via rotenone/antimycin A and 2-dg)
  • Energy Phenotype Test Kit (Agilent #103325-100)
    • mitochondrial respiration, glycolysis, and rapid determination of metabolic phenotype/switching (via oligomycin and FCCP)
  • Mitochondrial Fuel Flex Test Kit (Agilent #103260-100)
    • dependency, capacity, and flexibility of cells to oxidize three major mitochondrial fuels: glucose (pyruvate), glutamine (glutamate), and Long-chain fatty acids, in basal energetic state (using inhibitors: UK5099, BPTES and Etomoxir)
  • Mitochondrial Substrate Preference (Agilent #102720-100)
    • Respiration and substrate preference in responses to stress (via palmitate-BSA fatty acid oxidation substrate).
  • Glycolysis Stress Test Kit (Agilent #103020-100)
    • basal glycolysis and glycolysis capacity upon blockage of mitochondrial ATP production using respiratory modulators (oligomycin, glucose, and 2-dg)