Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)


  • Product name
    Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit
    See all Pyruvate dehydrogenase kits
  • Detection method
  • Sample type
    Cell culture extracts, Tissue Lysate, Purified mitochondria
  • Assay type
    Enzyme activity
  • Assay time
    3h 30m
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902) can be used to determine the activity of PDH in a human, bovine, mouse, or rat sample. The PDH enzyme is immunocaptured within the wells of the microplate and activity is determined by following the reduction of NAD+ to NADH, coupled to the reduction of a reporter dye to yield a colored reaction product with an increase in absorbance at 450 nm at room temperature. Included for performance of the activity assay are buffer, detergent, reagent mix, and a 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.

    This assay is optimized for use with whole tissue extract when the amount of total material available for assay is 20-100 µg or more. If using cell extract of cultured cells 1 mg of material is required due to the very low levels of enzyme and reduced levels of mitochondria in the extract.

    The PDH complex is relatively fragile and sensitive to detergent. Please follow the sample preparation steps provided in the protocol booklet. Other preparation methods may decrease PDH activity. Our Scientific Support team is happy to answer any questions about sample prep.

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Notes

    Store 20X Buffer, Detergent, and Microplate at 4°C.

    Store 5X Stabilizer at -20°C.

    Store 20X Reagent Mix, 100X Reagent Dye and 100X Coupler at -80°C.

  • Platform
    Microplate reader



  • Mitochondria, tissue extracts and whole cultured cell extracts all show linear relationships between signal and sample load at limiting concentrations. The rates shown were determined as change in OD over time, and these are best represented as change in milliOD per minute.
  • Example of microplate reader recorded data from bovine heart mitochondria (100 µg/well) (top trace) and 2-fold dilutions (stepwise lower traces) using Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
  • Schematic diagram showing the reaction involved in Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
  • Principle of Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)



This product has been referenced in:
  • Scheffler K  et al. 8-oxoguanine DNA glycosylase (Ogg1) controls hepatic gluconeogenesis. DNA Repair (Amst) 61:56-62 (2018). Read more (PubMed: 29207315) »
  • Corbet C  et al. Interruption of lactate uptake by inhibiting mitochondrial pyruvate transport unravels direct antitumor and radiosensitizing effects. Nat Commun 9:1208 (2018). Read more (PubMed: 29572438) »
See all 39 Publications for this product

Customer reviews and Q&As

Filter by Ratings

1-3 of 3 Abreviews

Cells were collected, counted and resuspended in PBS to adjust the sample protein concentration to 15 mg/mL. Then, samples were solubilized with Detergent (9:1), incubated on ice for 10 min and centrifuged at 1,000 g for 10 min at 4 ºC. The supernatants were diluted in Assay Buffer to the appropriated concentration within the linear working range for the assay, loaded to each well of the microplate and incubated at room temperature for 3 h. For the measurement, wells were emptied and 200 μL of Assay Solution were added. The absorbance
was measured at 450 nm using a kinetic program with readings every 25 sec for 30 min. PDH activity was normalized by protein content in the supernatant, determined by the bicinchoninic acid (BCA) assay.
It is important to notice that absorbance rapidly increases with time and therefore the time spent when adding the Assay Solution between wells is crucial.

Abcam user community

Verified customer

Submitted Oct 03 2018

The product was tested on mitochondria isolated from fetal sheep hearts. Activity was obtained with sheep heart mitochondria and the raw data ( starting from 12.5 ng/well to 400ng/well) is attached below.

Abcam user community

Verified customer

Submitted Jan 09 2018

Pyruvate dehydrogenase activity in HCT116 cells

Average Average 3/5 (Ease of Use)
We wanted to analyse the enzymatic activity of pyruvate dehydrogenase (PDH) in human HCT116 cells dependent on the glucose level within the media. Therefore we cultivated these cells in DMEM with high (4.5 g/l) and low (1 g/l) glucose for 48 h. They were harvested and lysed in PBS. We used whole cell lysates to purify proteins. The PDH activity was spectroscopically measured by a change in absorbance at 450 nm. The absorbance was measured every minute for 30 min and kinetics were analysed with GraFit. The changed absorbance per minute was plotted against protein mass used to load the plate (fig. A and B). The blank value was calculated from four blanks in each test.
Unfortunately we only obtained significant activities at 500 µg protein or higher (fig. A). Thus we repeated the test with higher protein concentrations (fig. B). But in both test, no substantial difference between cells cultivated in high or low glucose was observed.
For these assays we had the problem that the four blanks in each test had a high inter-plate variation. This may be due to our plate reader or to the plate delivered with the assay kit. Therefore you should always use technical replicates. Additionally we think that problems can easily appear if the protein quantification during the test is incorrect and accordingly the loading of the plate.
In general the assay kit is not easy to use but gives a quick method to analyse the PDH activity.

Mr. Christian Marx

Verified customer

Submitted Oct 04 2013

For licensing inquiries, please contact

Sign up