• Product name

    Glycolysis Assay [Extracellular acidification]
  • Detection method

  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based (quantitative)
  • Assay time

    1h 30m
  • Product overview

    Glycolysis Assay [Extracellular Acidification] (ab197244) is an easy mix-and-measure, 96 well fluorescence plate reader-based approach for the analysis of extracellular acidification (ECA/ECAR). As lactate production is the main contributor to this acidification, ab197244 is a convenient and informative measure of cellular glycolytic flux. Such measurements offer an important insight into the central role played by altered glycolytic activity in a wide array of physiological and pathophysiological processes, including cellular adaptation to hypoxia and ischemia, and the development and progression of tumorigenesis.

    The performance of Glycolysis Assay facilitates sensitive robust microtiter-plate based measurements, thereby overcoming many of the problems associated with the more cumbersome potentiometric pH approach. Rates of extracellular acidification are calculated from changes in fluorescence signal over time and, as the measurement is non-destructive and fully reversible (pH-sensitive reagent is not consumed), measurement of time-courses and multiple drug treatments are possible.

  • Notes

    Learn more about the full range of assays to measure glycolysis, oxygen consumption, fatty acid oxidation and metabolic flux in live cells.

    Or review the full metabolism assay guide for other assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress.

  • Platform

    Microplate reader



  • Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 µM] and FCCP [2.5 µM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).

  • Typical Lifetime profile of Glycolysis Assay for adherent cells, treated with typical control compounds, including Oxamic acid recommended as a negative control. The effect of Glucose Oxidase as a positive signal control is illustrated schematically.

  • Excitation and Emission spectra of Glycolysis Assay. Left panel shows normalized excitation (Ex 340 – 410nm; Peak 360-380nm). Right panel shows emission maxima (Em 590, 615 and 690nm) fold increase between pH6.0 and pH7.5.



This product has been referenced in:

  • Sun Y  et al. Inhibition of mitochondrial respiration overcomes hepatocellular carcinoma chemoresistance. Biochem Biophys Res Commun 508:626-632 (2019). Read more (PubMed: 30522865) »
  • Valentín-Guillama G  et al. HIV-1 Envelope Protein gp120 Promotes Proliferation and the Activation of Glycolysis in Glioma Cell. Cancers (Basel) 10:N/A (2018). Read more (PubMed: 30200472) »
See all 4 Publications for this product

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