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An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes

An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes

DOI:
10.3791/54918-v

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08:40 min

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November 21, 2016

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, , , ,
1Laboratory of Mitochondrial Biology and Metabolism, National Heart, Lung, and Blood Institute,National Institutes of Health, 2Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases,National Institutes of Health, 3Laboratory of Immunology, National Institute of Allergy and Infectious Diseases,National Institutes of Health

Capitoli

  • 00:05Titolo
  • 01:04Tissue Processing
  • 03:28Magnetic B Cell and Naïve CD4+ T Cell Separation
  • 04:43Extracellular Flux Assay
  • 06:09Results: Representative Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) Analyses
  • 07:48Conclusion

Summary

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The study of metabolism is becoming increasingly relevant to immunological research. Here, we present an optimized method for measuring glycolysis and mitochondrial respiration in mouse splenocytes, and T and B lymphocytes.

Tags

Keywords: GlycolysisMitochondrial RespirationLymphocytesExtracellular Flux AssayImmunometabolismMetabolic ProfilesNon-adherent CellsCell FunctionalityCell ViabilityCell StrainerACK Red Blood Cell Lysis BufferMS BufferCell SuspensionCell PelletCentrifugation

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