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Cellular metabolism assays

Related

  • Browse metabolism assays
    • ROS, oxidative stress, and antioxidants
      • Assays for mitochondrial research
        • Metabolism research resources

          Simple assays and biochemicals for your metabolism research

          ​Metabolism is a complex process that lies at the core of biology. Changes to metabolism are involved in a huge range of outcomes, from cancer to neurodegeneration, and more.

          Make your metabolism research less complex, with easy-to-use assay kits. Analyze live cells, lysates, and biofluids, with readout on your plate reader, microscope, or flow cytometer.

          Download our  metabolism assays guide or review our assays below

          Contents

          • Metabolites and enzymes of sugar, lipid, alcohol, and amino acid metabolism
          • Metabolic activity analysis
          • ATP, NADH, and associated molecules

          Metabolites and enzymes of sugar, lipid, alcohol, and amino acid metabolism

          Find your ideal assay to quantify metabolite levels or enzyme activity. Our assay kits allow you to analyze the pathways of metabolism in depth in live cells, lysates and biofluids with easy analysis on your plate reader. 

          Browse assays for:

          • Lipid metabolism
          • ​Sugar metabolism
          • Amino acid metabolism 
          • Coenzymes and cofactors​​​
          • ​Intermediary metabolism (including glycolysis and the citric acid cycle)​
          Or search for your analyte or enzyme of interest for areas not covered above.

          Metabolic activity analysis: oxygen consumption, glycolysis, and fatty acid metabolism

          Microplate-based assays to measure O2 consumption or glycolytic flux in live cells without the need for specialized equipment.
          A fluorescent-based approach for direct real-time analysis of cellular respiration and glycolytic flux.

          • Assays are based on oxygen- and pH-sensing fluorophores​
          • Reactions are non-destructive and fully reversible, allowing measurement of time courses and drug treatments
          • Assays do not require specialized equipment or probes; signal is measured in a standard fluorescence or TR-F plate reader​
          ​

          Assay

          What it measures

          How it works

          Assay kits

          Extracellular Oxygen Consumption (OCR) Assay

          O2 consumption rate

          As cell respiration lowers O2 concentration, dye fluorescence increases

          ab197243, ab197242


          Intracellular Oxygen Assay

          Intra-cellular O2 levels

          Dye fluorescence is inversely proportional to oxygen concentration

          ab197245

          Glycolysis Assay (ECAR)

          Extracellular acidification (glycolysis) rate

          Lactate causes extracellular acidification, dye fluorescence increases

          Core assay: ab197244
          Glycolysis stress test kits: ab222945 and ab222946

          Fatty Acid Oxidation Assay

          O2 consumption rate on blocking of sugar metabolism

          As fatty acid oxidation lowers O2 concentration, dye fluorescence increases

          Companion kit to OCR assay: ab217602
          Complete kit: ab222944


          ​

          Case study: Measurement of mitochondrial metabolism in cultured cells

          Oxygen consumption rate and extracellular acidification assays can be used together to build a metabolic picture of response to drug treatment. 

          We measured oxidative phosphorylation and anaerobic glycolytic flux in the human hepato-carcinoma cell line HepG2 (Figure 1). Cells were treated with FCCP (an OXPHOS uncoupler) and antimycin A (a complex III inhibitor). Cellular energy production was monitored by measuring ATP content.​​

          • Oxygen consumption was measured using the Extracellular Oxygen Consumption Assay under a mineral oil seal to prevent the back diffusion of ambient oxygen.
          • Glycolytic flux was measured with the Glycolysis Assay [Extracellular Acidification]. Measurements were made using unsealed samples to avoid CO2​ contribution to extracellular acidification​.
          • ATP concentration was measured with Luminescent ATP Detection Assay. ATP was quantified using unsealed samples after ECA measurements.​
          ab113849 ATP detection 472px

          ​

          Figure 1: HepG2 cells (seeded at 6.5 x 104 cells/well) were treated with 1 µM antimycin A and 2.5 µM FCCP. Oxygen consumption (white column), extracellular acidification rate (black column) and ATP concentration (stripped column) data are shown as percentage of untreated control. All measurements were performed on a FLUOstar Omega (BMG Labtech).

          As expected, complex III inhibition by antimycin A reduced oxygen consumption rate to undetectable levels, while FCCP treatment significantly increased oxygen consumption. Both antimycin A and FCCP treatment caused a dramatic increase in extracellular acidification due to the increase in lactate production. Cellular ATP concentration was virtually unchanged in treated cells as energy production is maintained.

          In conclusion, we have described how to study changes to glycolytic and oxidative metabolism in response to mitochondrial modulators in real-time with a simple microplate-based assay.​

          Assays for ATP, NADH, and associated molecules​

          ​ATP assays are either based on the extraction of ATP from cells followed by light production with ATP-dependent luciferase, or the ATP-dependent phosphorylation of glycerol (or other substrates) to generate a detectable product.

          Assay

          Read out

          Notes

          Assay kits

          ATP

          Plate reader

          No-wash assay.

          ab113849

          ADP/ATP

          No-wash assay. Same method as ATP assay. After ATP analysis, ADP is converted to ATP and detected.

          ab65313

          ATP

          Used with cell lysates.

          Not as sensitive as luminescence assays.

          ab83355

          Phosphate


          ab65622, ab219938 ab102508

          Pyrophosphate


          ab112155 ab179836


          NADH, NAD, NADHP and NADPH

          Assay

          Readout

          Assay kits

          NADH

          Plate reader

          ab186030

          NAD/NADH

          ab65348, ab176723, ab186032

          NADP / NADPH

          ab186033, ab65349, ab176724

          NADPH

          ab186031




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