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Glycolysis Assay [Extracellular acidification] (ab197244)

  • Datasheet
  • SDS
  • Protocol Booklet
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Simultaneous quantification of mitochondrial respiration and glycolytic flux
  • Typical lifetime profile
  • Excitation and emission spectra

Key features and details

  • Assay type: Cell-based (quantitative)
  • Detection method: Fluorescent
  • Platform: Microplate reader
  • Assay time: 1 hr 30 min
  • Sample type: Adherent cells, Suspension cells

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Overview

  • Product name

    Glycolysis Assay [Extracellular acidification]
  • Detection method

    Fluorescent
  • 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

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 96 tests 4 x 96 tests
    Glycolysis Assay Reagent (lyophilized) 1 vial 4 vials
    Respiration Buffer 1 tablet 4 tablets
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Intermediary Metabolism Kits
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Oxidative Stress Assay Kits
    • Oxidative Stress

Associated products

  • Related Products

    • Extracellular Oxygen Consumption Reagent (ab197242)
    • Extracellular Oxygen Consumption Assay (ab197243)
    • Intracellular Oxygen Concentration Assay (ab197245)
    • Glycolysis Stress Test Companion Assay (ab222945)

Images

  • Simultaneous quantification of mitochondrial respiration and glycolytic flux
    Simultaneous quantification of mitochondrial respiration and glycolytic flux

    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
    Typical lifetime profile

    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
    Excitation and emission spectra

    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.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (23)

Publishing research using ab197244? Please let us know so that we can cite the reference in this datasheet.

ab197244 has been referenced in 23 publications.

  • Horikawa M  et al. Dual roles of AMAP1 in the transcriptional regulation and intracellular trafficking of carbonic anhydrase IX. Transl Oncol 15:101258 (2022). PubMed: 34742153
  • Lang L  et al. Tumor suppressive role of microRNA-4731-5p in breast cancer through reduction of PAICS-induced FAK phosphorylation. Cell Death Discov 8:154 (2022). PubMed: 35379785
  • Fang Y  et al. LAMC1-mediated preadipocytes differentiation promoted peritoneum pre-metastatic niche formation and gastric cancer metastasis. Int J Biol Sci 18:3082-3101 (2022). PubMed: 35541892
  • Zhang J  et al. TMPRSS12 Functions in Meiosis and Spermiogenesis and Is Required for Male Fertility in Mice. Front Cell Dev Biol 10:757042 (2022). PubMed: 35547804
  • You JH  et al. PGRMC1-dependent lipophagy promotes ferroptosis in paclitaxel-tolerant persister cancer cells. J Exp Clin Cancer Res 40:350 (2021). PubMed: 34749765
View all Publications for this product

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