Pyruvate Measurement
Pyruvate Measurement - Related Videos
02:29
Pyruvate Assay for Estimation of Pyruvate Levels in Cellular Fractions of Worms
0 Views •
2025
This video describes a colorimetric assay for quantitatively measuring pyruvate in the cellular fractions of Caenorhabditis elegans. Utilizing the hydrogen peroxide produced from the enzymatic reaction of pyruvate oxidase with cellular pyruvate, a chromogenic dye is oxidized to produce a colored product, measured using colorimetry to determine the pyruvate concentration.
01:15
Pyruvate Oxidation
0 Views •
2019
After glycolysis, the charged pyruvate molecules enter the mitochondria via active transport and undergo three enzymatic reactions. These reactions ensure that pyruvate can enter the next metabolic pathway so that energy stored in the pyruvate molecules can be harnessed by the cells. First, the enzyme pyruvate dehydrogenase removes the carboxyl group from pyruvate and releases it as carbon dioxide. The stripped molecule is then oxidized and releases electrons, which are then picked up by NAD+...
01:20
Fates of Pyruvate
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2025
Pyruvate is the end product of glycolysis, where glucose is oxidized to pyruvate, simultaneously reducing NAD+ to NADH. Two molecules of ATP are also produced by substrate-level phosphorylation. In aerobic organisms, pyruvate is metabolized via the citric acid cycle to produce reduced coenzymes NADH and FADH2. These coenzymes are then oxidized in the electron transport chain to produce ATP and, in the process, regenerate the NAD+ and FAD. As seen in some cell types and organisms, fermentation...
![Measuring the Spin-Lattice Relaxation Magnetic Field Dependence of Hyperpolarized [1-<sup>13</sup>C]pyruvate](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F59399.jpg%3Fv%3D2026-09-01%252018%253A12%253A11&w=750&q=75)
11:57
Measuring the Spin-Lattice Relaxation Magnetic Field Dependence of Hyperpolarized [1-13C]pyruvate
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Cited by 2 •
2019
We present a protocol to measure the magnetic field dependence of the spin-lattice relaxation time of 13C-enriched compounds, hyperpolarized by means of dynamic nuclear polarization, using fast field-cycled relaxometry. Specifically, we have demonstrated this with [1-13C]pyruvate, but the protocol could be extended to other hyperpolarized substrates.
09:35
Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans
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Cited by 2 •
2018
We describe a modified small-scale extraction and colorimetric assays of lactate and pyruvate in the nematode C. elegans. When utilizing commercial assay kits, the technical development of their sensitivity and accuracy is important. Protein precipitation in extraction is the most critical step for the quantitative determination of intracellular metabolites.
FAQs
The assay couples pyruvate conversion to a change in NADH. Lactate dehydrogenase uses pyruvate to form lactate while NADH becomes NAD+. Because NADH contributes to absorbance at 340 nm, its loss produces a declining optical signal. The magnitude of this decrease provides a quantitative basis for comparing pyruvate levels among biological samples.
Pyruvate does not necessarily remain unchanged after a sample is collected. Cells or tissue-associated processes can rapidly consume it or convert it into other compounds, altering the concentration that the assay measures. Rapid processing helps preserve the sample’s metabolic state at collection, improving the relevance of the result to the original biological condition.
Pyruvate levels can help indicate how cellular energy metabolism is balanced between glycolysis and mitochondrial oxidation. When interpreted with an appropriate experimental design, the measurement can contribute to studies of metabolic flux, meaning movement through metabolic pathways. It therefore links a biochemical readout with broader changes in cellular energy handling.
A basic workflow includes collecting the biological sample, processing it rapidly, exposing it to the lactate dehydrogenase reaction, and monitoring absorbance at 340 nm. The observed decrease in absorbance is then used to determine the pyruvate amount. Consistent handling across samples is important because post-collection conversion can affect comparisons.
Researchers may select this measurement when examining tissue function, disease-associated metabolic changes, or the effects of drugs and altered culture conditions. It can show whether an intervention is associated with a changed pyruvate state, while also supporting investigation of how cellular energy metabolism responds to experimental or physiological conditions.
The technique provides a quantitative endpoint for comparing samples exposed to different conditions. Differences in the measured signal may help investigators evaluate changes linked to treatment, disease state, or culture environment. In tissue studies, the result can be considered alongside tissue function, whereas in cellular experiments it can support analysis of altered metabolic behavior.