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AB109902

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit

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(4 Reviews)

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(107 Publications)

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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Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902)
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Supplier Data

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)
  • FuncS

Supplier Data

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).

Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902)
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Supplier Data

Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902)

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

Schematic Diagram - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (AB109902)
  • Schematic Diagram

Supplier Data

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).

Key facts

Detection method

Colorimetric

Sample types

Tissue Lysate, Cell culture extracts, Purified mitochondria

Reacts with

Mouse, Rat, Cow, Human

Assay type

Enzyme activity

Assay time

3h 30m

Assay Platform

Microplate reader

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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 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.

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What's included?

{ "values": { "96Test": { "sellingSize": "96 Test", "publicAssetCode":"ab109902-96Test", "assetComponentDetails": [ { "size":"1 x 15 mL", "name":"20X Buffer", "number":"AB109902-CMP01", "productcode":"" }, { "size":"1 x 13 mL", "name":"5X Stabilizer", "number":"AB109902-CMP04", "productcode":"" }, { "size":"2 x 600 µL", "name":"20X Reagent Mix", "number":"AB109902-CMP02", "productcode":"" }, { "size":"1 x 1 Unit", "name":"96-well Microplate (12 strips)", "number":"AB109902-CMP05", "productcode":"" }, { "size":"2 x 1 mL", "name":"Detergent", "number":"AB109902-CMP07", "productcode":"" }, { "size":"1 x 250 µL", "name":"Coupler", "number":"AB109902-CMP03", "productcode":"" }, { "size":"1 x 250 µL", "name":"Reagent Dye", "number":"AB109902-CMP06", "productcode":"" } ] } } }
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Properties and storage information

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

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

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.

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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Product protocols

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Target data

See full target information Pyruvate Dehydrogenase
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Publications (107)

Recent publications for all applications. Explore the full list and refine your search

Nature communications 16:3177 PubMed40180922

2025

Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation.

Applications

Unspecified application

Species

Unspecified reactive species

Ning Zhang,Linmao Sun,Shuo Zhou,Changyong Ji,Tianming Cui,Qi Chu,Jiareng Ye,Shuhang Liang,Kun Ma,Yufeng Liu,Xianying Li,Xinyu Guo,Weizhi Zhang,Xuetian Gu,Cheng Cheng,Qingrui Zha,Shengwei Tao,Yunguang Zhang,Junhui Chu,Chenghui Wu,Yuchen Zhang,Jiabei Wang,Yao Liu,Lianxin Liu

International journal of molecular sciences 26: PubMed39796278

2025

Glutaminase-2 Expression Induces Metabolic Changes and Regulates Pyruvate Dehydrogenase Activity in Glioblastoma Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Juan De Los Santos-Jiménez,José A Campos-Sandoval,Tracy Rosales,Bookyung Ko,Francisco J Alonso,Javier Márquez,Ralph J DeBerardinis,José M Matés

Nature communications 15:9945 PubMed39550366

2024

NOTCH1 mitochondria localization during heart development promotes mitochondrial metabolism and the endothelial-to-mesenchymal transition in mice.

Applications

Unspecified application

Species

Unspecified reactive species

Jie Wang,Rui Zhao,Sha Xu,Xiang-Yu Zhou,Ke Cai,Yu-Ling Chen,Ze-Yu Zhou,Xin Sun,Yan Shi,Feng Wang,Yong-Hao Gui,Hui Tao,Jian-Yuan Zhao

Basic & clinical pharmacology & toxicology 136:e14102 PubMed39501987

2024

Dichloroacetate, a pyruvate dehydrogenase activator, alleviates high-fat-induced impairment of myogenic differentiation in skeletal muscles.

Applications

Unspecified application

Species

Unspecified reactive species

Chuang-Yen Huang,I-Shan Han,Po-Shiuan Hsieh,Min-Chien Tsai,Hung-Che Chien

Frontiers in molecular biosciences 11:1460987 PubMed39297074

2024

Recent progress of methods for cuproptosis detection.

Applications

Unspecified application

Species

Unspecified reactive species

Ligang Zhang,Ruiting Deng,Raoqing Guo,Yawen Jiang,Yichen Guan,Caiyue Chen,Wudi Zhao,Guobin Huang,Lian Liu,Hongli Du,Dongsheng Tang

Cell death & disease 15:588 PubMed39138149

2024

Dihydrolipoamide dehydrogenase (DLD) is a novel molecular target of bortezomib.

Applications

Unspecified application

Species

Unspecified reactive species

Yu Feng,Hongmei Luo,Jingcao Huang,Yue Zhang,Jingjing Wen,Linfeng Li,Ziyue Mi,Qianwen Gao,Siyao He,Xiang Liu,Xinyu Zhai,Xin Wang,Li Zhang,Ting Niu,Yuhuan Zheng

eLife 13: PubMed38757931

2024

PI3K/HSCB axis facilitates FOG1 nuclear translocation to promote erythropoiesis and megakaryopoiesis.

Applications

Unspecified application

Species

Unspecified reactive species

Gang Liu,Yunxuan Hou,Xin Jin,Yixue Zhang,Chaoyue Sun,Chengquan Huang,Yujie Ren,Jianmin Gao,Xiuli Wang,Xiumei Jiang

Cell 187:2359-2374.e18 PubMed38653240

2024

BCAA-nitrogen flux in brown fat controls metabolic health independent of thermogenesis.

Applications

Unspecified application

Species

Unspecified reactive species

Anthony R P Verkerke,Dandan Wang,Naofumi Yoshida,Zachary H Taxin,Xu Shi,Shuning Zheng,Yuka Li,Christopher Auger,Satoshi Oikawa,Jin-Seon Yook,Melia Granath-Panelo,Wentao He,Guo-Fang Zhang,Mami Matsushita,Masayuki Saito,Robert E Gerszten,Evanna L Mills,Alexander S Banks,Yasushi Ishihama,Phillip J White,Robert W McGarrah,Takeshi Yoneshiro,Shingo Kajimura

Cell reports 43:113854 PubMed38412099

2024

Flvcr1a deficiency promotes heme-based energy metabolism dysfunction in skeletal muscle.

Applications

Unspecified application

Species

Unspecified reactive species

Miriam Mistretta,Veronica Fiorito,Anna Lucia Allocco,Giorgia Ammirata,Myriam Y Hsu,Sabrina Digiovanni,Marzia Belicchi,Laura Napoli,Michela Ripolone,Elena Trombetta,PierLuigi Mauri,Andrea Farini,Mirella Meregalli,Chiara Villa,Paolo Ettore Porporato,Barbara Miniscalco,Simonetta Geninatti Crich,Chiara Riganti,Yvan Torrente,Emanuela Tolosano

Cell research 34:13-30 PubMed38163844

2024

Hypoxia induces mitochondrial protein lactylation to limit oxidative phosphorylation.

Applications

Unspecified application

Species

Unspecified reactive species

Yunzi 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 Zhao
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