The oxygen consumption rate was analyzed using an XFp Extracellular Flux Analyzer (Agilent, CA, USA). iPSC-derived neurons were plated on XFp microplates (Agilent, CA, USA) at a density of 70,000 per well and grown in N2 medium for 48 h before the experiment. Measurement of the neuronal oxidative consumption rate was performed in a freshly prepared medium consisting of phenol-free DMEM, 1 mM sodium pyruvate, 2 mM glutamine, and 10 mM glucose with the pH adjusted to 7.4. Mitochondrial function was evaluated at baseline levels and after the subsequent injection of 10 μM oligomycin, 10 μM carbonyl cyanide p-trifluoromethoxyphenylhydrazone, and 1 μM antimycin A/1 μM rotenone (all Sigma-Aldrich, MO, USA). Three measurements, lasting 5 min each, were taken for each condition. The measured values were normalized on protein concentration measured by BCA.
Xfp microplates
Manufactured by Agilent Technologies
Sourced in United States
The XFp microplates are a line of consumable laboratory equipment designed for use with Agilent's Seahorse XFp Analyzer. The plates are intended for performing extracellular flux analysis, which measures the oxygen consumption rate and extracellular acidification rate of cells. The microplates are made of a specialized material and feature a multi-well format to accommodate multiple samples or conditions simultaneously.
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Lab products found in correlation
2 protocols using xfp microplates
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Neuronal Metabolic Profiling with XFp
2
Neuronal Metabolic Profiling with XFp
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The oxygen consumption rate was analyzed using an XFp Extracellular Flux Analyzer (Agilent, CA, USA). iPSC-derived neurons were plated on XFp microplates (Agilent, CA, USA) at a density of 70,000 per well and grown in N2 medium for 48 h before the experiment. Measurement of the neuronal oxidative consumption rate was performed in a freshly prepared medium consisting of phenol-free DMEM, 1 mM sodium pyruvate, 2 mM glutamine, and 10 mM glucose with the pH adjusted to 7.4. Mitochondrial function was evaluated at baseline levels and after the subsequent injection of 10 μM oligomycin, 10 μM carbonyl cyanide p-trifluoromethoxyphenylhydrazone, and 1 μM antimycin A/1 μM rotenone (all Sigma-Aldrich, MO, USA). Three measurements, lasting 5 min each, were taken for each condition. The measured values were normalized on protein concentration measured by BCA.
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