Determining Maximum Glycolytic Capacity Using Extracellular Flux Measurements

PLoS One. 2016 Mar 31;11(3):e0152016. doi: 10.1371/journal.pone.0152016. eCollection 2016.


Measurements of glycolytic rate and maximum glycolytic capacity using extracellular flux analysis can give crucial information about cell status and phenotype during normal operation, development of pathology, differentiation, and malignant transformation. They are also of great use when assessing the effects of chemical or drug treatments. Here, we experimentally define maximum glycolytic capacity, demonstrate how it differs from glycolytic rate, and provide a protocol for determining the basal glycolytic rate and maximum glycolytic capacity in cells using extracellular flux measurements. The results illustrate the power of extracellular flux analysis to describe the energetics of adherent cells in culture in a fully quantitative way.

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Animals
  • Biochemistry / methods
  • Cell Line
  • Cell Respiration
  • Fibroblasts / metabolism*
  • Glycolysis*
  • HEK293 Cells
  • Humans
  • Mice
  • Myoblasts / metabolism*
  • Oxidative Phosphorylation
  • Sodium-Potassium-Exchanging ATPase / metabolism


  • Adenosine Triphosphate
  • Sodium-Potassium-Exchanging ATPase

Grant support

The authors have no support or funding to report.