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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
Metabolites and enzymes of sugar, lipid, alcohol, and amino acid metabolism
What it measures
How it works
Extracellular Oxygen Consumption (OCR) Assay
O2 consumption rate
As cell respiration lowers O2 concentration, dye fluorescence increases
Intracellular Oxygen Assay
Intra-cellular O2 levels
Dye fluorescence is inversely proportional to oxygen concentration
Glycolysis Assay (ECAR)
Extracellular acidification (glycolysis) rate
Lactate causes extracellular acidification, dye fluorescence increases
Fatty Acid Oxidation Assay
O2 consumption rate on blocking of sugar metabolism
As fatty acid oxidation lowers O2 concentration, dye fluorescence increases
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.
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.
No-wash assay. Same method as ATP assay. After ATP analysis, ADP is converted to ATP and detected.
Used with cell lysates.
Not as sensitive as luminescence assays.
NADH, NAD, NADHP and NADPH
NADP / NADPH