Cellular metabolism assays

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

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:

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​
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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


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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.​​

ab113849 ATP detection 472px

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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