Skip to main content

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. Readout on any colorimetric (450 nm) plate reader.

- Cited in > 100 publications


Images

Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902), expandable thumbnail
  • Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902), expandable thumbnail
  • Schematic Diagram - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902), expandable thumbnail
  • Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric
Sample types
Tissue Lysate, Cell culture extracts, Purified mitochondria
Assay type
Enzyme activity
Reactive species
Mouse, Rat, Cow, Human
Assay time
3h 30m

What's included?

96 Test
Components
20X Buffer
1 x 15 mL
20X Reagent Mix
2 x 600 µL
5X Stabilizer
1 x 13 mL
96-well Microplate (12 strips)
1 x 1 Unit
Coupler
1 x 250 µL
Detergent
2 x 1 mL
Reagent Dye
1 x 250 µL

Recommended products

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. Readout on any colorimetric (450 nm) plate reader.

- Cited in > 100 publications

Key facts

Detection method
Colorimetric
Sample types
Tissue Lysate, Cell culture extracts, Purified mitochondria
Assay type
Enzyme activity
Reactive species
Mouse, Rat, Cow, Human
Assay time
3h 30m
Assay Platform
Microplate reader

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
Multi
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols

Notes

The Pyruvate Dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit works by immunocapturing the PDH enzyme in microplate wells and measuring its activity through the reduction of NAD+ to NADH, which is linked to a color change detected at 450 nm.

How the assay works

In Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit protocol, 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.

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit protocol summary:

- Add samples, standards and positive control to wells
- Incubate for 3 hours at room temperature
- Add 200 µL of Assay Solution to each well
- Measure Optical Density (OD450 nm) in a kinetic mode at RT for 30 minutes

How other researchers are using

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit has been used in a variety of sample type including:
Mouse cardiomyocytes 1
Mouse muscle tissue samples 2
Human iPSC-derived cardiomyocytes 3
References: 

1-Mao Y et al. 2023
2-Brenmoehl J et al. 2023
3-Srivastava S et al. 2023

Related and recommended products

See an alternative kit to quantify Pyruvate dehydrogenase (PDH) Enzyme Activity:
- Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit ab109882

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

Store 5X Stabilizer, 20X Reagent Mix, 100X Reagent Dye and 100X Coupler at -80°C.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Pyruvate dehydrogenase (PDH) is an enzyme complex also known as the pyruvate dehydrogenase complex (PDC) or PDH complex. It is a multi-enzyme structure with a mass on the order of megadaltons and plays an important role in cellular energy metabolism. Situated in the mitochondrial matrix pyruvate dehydrogenase converts pyruvate into acetyl-CoA through oxidative decarboxylation. This conversion releases one molecule of CO2 and reduces NAD+ to NADH. The complex includes three core enzymes: E1 (pyruvate dehydrogenase) E2 (dihydrolipoamide acetyltransferase) and E3 (dihydrolipoamide dehydrogenase).

Biological function summary

The actions of pyruvate dehydrogenase serve as a bridge between glycolysis and the tricarboxylic acid (TCA) cycle. Pyruvate derived from glucose is transformed into acetyl-CoA before entering the TCA cycle for further energy extraction. The PDH complex ensures efficient energy production by tightly regulating the flow of carbon into the TCA cycle. Regulation occurs through phosphorylation by specific PDH kinases which inactivate E1. This mechanism integrates signals from energy status and substrates availability modulating the carbohydrate metabolism.

Pathways

Pyruvate dehydrogenase is a central player in cellular respiration and energy metabolism. It connects glycolytic pathways with the TCA cycle facilitating energy conversion in eukaryotic cells. Key related proteins involve pyruvate kinase (which generates pyruvate) and citrate synthase (which uses acetyl-CoA) ensuring synchronized activity between upstream and downstream metabolic processes. The proper function of PDH activity is necessary for maintaining the metabolic flow with the PDH complex serving a gating role in the energy pathways.

Associated diseases and disorders

Pyruvate dehydrogenase deficiency results in metabolic challenges as the inability to convert pyruvate efficiently causes an increase in lactate levels. This condition results in lactic acidosis and severe neurological dysfunction. Furthermore alterations in PDH activity are observed in various forms of cancer as cancer cells often rely on aerobic glycolysis (Warburg effect) rather than complete oxidation of glucose. In this context the PDH protein interacts with oncogenic pathways highlighting its role in tumor metabolism and potential therapeutic targeting.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902), expandable thumbnail

    Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

    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.

  • Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902), expandable thumbnail

    Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

    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 - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902), expandable thumbnail

    Schematic Diagram - 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).

  • Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902), expandable thumbnail

    Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

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

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com