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
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- Cell research 34:13-302024Hypoxia induces mitochondrial protein lactylation to limit oxidative phosphorylation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYunzi Mao,Jiaojiao Zhang,Qian Zhou,Xiadi He,Zhifang Zheng,Yun Wei,Kaiqiang Zhou,Yan Lin,Haowen Yu,Haihui Zhang,Yineng Zhou,Pengcheng Lin,Baixing Wu,Yiyuan Yuan,Jianyuan Zhao,Wei Xu,Shimin ZhaoPubMed 38163844
- Cells 12:2023Metabolic Pathway Modeling in Muscle of Male Marathon Mice (DUhTP) and Controls (DUC)-A Possible Role of Lactate Dehydrogenase in Metabolic Flexibility.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJulia Brenmoehl,Elli Brosig,Nares Trakooljul,Christina Walz,Daniela Ohde,Antonia Noce,Michael Walz,Martina Langhammer,Stefan Petkov,Monika Röntgen,Steffen Maak,Christina E Galuska,Beate Fuchs,Björn Kuhla,Siriluck Ponsuksili,Klaus Wimmers,Andreas HoeflichPubMed 37566003
- Nature communications 14:43602023Nuclear translocation of mitochondrial dehydrogenases as an adaptive cardioprotective mechanism.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShubhi Srivastava,Priyanka Gajwani,Jordan Jousma,Hiroe Miyamoto,Youjeong Kwon,Arundhati Jana,Peter T Toth,Gege Yan,Sang-Ging Ong,Jalees RehmanPubMed 37468519
- Nature metabolism 5:1290-13022023Fatty acid oxidation organizes mitochondrial supercomplexes to sustain astrocytic ROS and cognition.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBrenda Morant-Ferrando,Daniel Jimenez-Blasco,Paula Alonso-Batan,Jesús Agulla,Rebeca Lapresa,Dario Garcia-Rodriguez,Sara Yunta-Sanchez,Irene Lopez-Fabuel,Emilio Fernandez,Peter Carmeliet,Angeles Almeida,Marina Garcia-Macia,Juan P BolañosPubMed 37460843
- Communications medicine 3:712023Imaging immunomodulatory treatment responses in a multiple sclerosis mouse model using hyperpolarized C metabolic MRI.Applications:Unspecified applicationReactive species:Unspecified reactive speciesCaroline Guglielmetti,Christian Cordano,Chloé Najac,Ari J Green,Myriam M ChaumeilPubMed 37217574
- Acta pharmaceutica Sinica. B 13:1145-11632023Targeting metabolic vulnerability in mitochondria conquers MEK inhibitor resistance in -mutant lung cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJuanjuan Feng,Zhengke Lian,Xinting Xia,Yue Lu,Kewen Hu,Yunpeng Zhang,Yanan Liu,Longmiao Hu,Kun Yuan,Zhenliang Sun,Xiufeng PangPubMed 36970205
- Nature communications 14:16662023Integrative proteogenomic characterization of early esophageal cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLingling Li,Dongxian Jiang,Qiao Zhang,Hui Liu,Fujiang Xu,Chunmei Guo,Zhaoyu Qin,Haixing Wang,Jinwen Feng,Yang Liu,Weijie Chen,Xue Zhang,Lin Bai,Sha Tian,Subei Tan,Chen Xu,Qi Song,Yalan Liu,Yunshi Zhong,Tianyin Chen,Pinghong Zhou,Jian-Yuan Zhao,Yingyong Hou,Chen DingPubMed 36966136
- Nutrients 15:2023Low-Carbohydrate Diet Modulates Glucose-Lipid Utilization in Skeletal Muscle of Diabetic Mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYan Li,Zi Yang,Yu Wang,Mingcong Fan,Chenzhipeng Nie,Lamei Xue,Li Wang,Haifeng QianPubMed 36986243
- Nature metabolism 4:693-7102022Cancer-associated fibroblasts require proline synthesis by PYCR1 for the deposition of pro-tumorigenic extracellular matrix.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmily J Kay,Karla Paterson,Carla Riera-Domingo,David Sumpton,J Henry M Däbritz,Saverio Tardito,Claudia Boldrini,Juan R Hernandez-Fernaud,Dimitris Athineos,Sandeep Dhayade,Ekaterina Stepanova,Enio Gjerga,Lisa J Neilson,Sergio Lilla,Ann Hedley,Grigorios Koulouras,Grace McGregor,Craig Jamieson,Radia Marie Johnson,Morag Park,Kristina Kirschner,Crispin Miller,Jurre J Kamphorst,Fabricio Loayza-Puch,Julio Saez-Rodriguez,Massimiliano Mazzone,Karen Blyth,Michele Zagnoni,Sara ZanivanPubMed 35760868
- Nature communications 13:21052022Insulin action and resistance are dependent on a GSK3β-FBXW7-ERRα transcriptional axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHui Xia,Charlotte Scholtes,Catherine R Dufour,Carlo Ouellet,Majid Ghahremani,Vincent GiguèrePubMed 35440636
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?
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.
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4 product images
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)
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)
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)
Principle of Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
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