Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit is an immunoassay kit for the detection of Pyruvate dehydrogenase (PDH) Enzyme Activity in Mouse, Rat, Cow, Human, Pig in Cell culture extracts, Tissue samples.
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- Orphanet journal of rare diseases 18:802023Alpha-lipoic acid supplementation corrects pathological alterations in cellular models of pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMarta Talaverón-Rey,Mónica Álvarez-Córdoba,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,David Gómez-Fernández,Ana Romero-González,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Paula Cilleros-Holgado,Diana Reche-López,Rocío Piñero-Pérez,José A Sánchez-AlcázarPubMed 37046296
- International journal of molecular sciences 23:2022A New Role of NAP1L1 in Megakaryocytes and Human Platelets.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMartin Freitag,Hansjörg SchwertzPubMed 36499021
- Orphanet journal of rare diseases 17:3112022Therapeutic approach with commercial supplements for pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMónica Álvarez-Córdoba,Diana Reche-López,Paula Cilleros-Holgado,Marta Talaverón-Rey,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Rocío Piñero-Pérez,José A Sánchez-AlcázarPubMed 35945593
- Orphanet journal of rare diseases 16:2012021Down regulation of the expression of mitochondrial phosphopantetheinyl-proteins in pantothenate kinase-associated neurodegeneration: pathophysiological consequences and therapeutic perspectives.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMónica Álvarez-Córdoba,Marta Talaverón-Rey,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Joaquín J Salas,José A Sánchez-AlcázarPubMed 33952316
- Cell reports 33:1083332020Metabolic Reprogramming Induces Germinal Center B Cell Differentiation through Bcl6 Locus Remodeling.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKei Haniuda,Saori Fukao,Daisuke KitamuraPubMed 33147467
- Proceedings of the National Academy of Sciences of 116:25389-253912019Repurposing dichloroacetate for the treatment of women with endometriosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAndrew W Horne et. al.PubMed 31792175
- Frontiers in cardiovascular medicine 5:1952019Biventricular Increases in Mitochondrial Fission Mediator (MiD51) and Proglycolytic Pyruvate Kinase (PKM2) Isoform in Experimental Group 2 Pulmonary Hypertension-Novel Mitochondrial Abnormalities.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPing Yu Xiong,Lian Tian,Kimberly J Dunham-Snary,Kuang-Hueih Chen,Jeffrey D Mewburn,Monica Neuber-Hess,Ashley Martin,Asish Dasgupta,Francois Potus,Stephen L ArcherPubMed 30740395
- JCI insight 4:2019The antianginal ranolazine mitigates obesity-induced nonalcoholic fatty liver disease and increases hepatic pyruvate dehydrogenase activity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRami Al Batran et. al.PubMed 30626749
- International journal of molecular sciences 19:2018Transcriptomic Signature of Right Ventricular Failure in Experimental Pulmonary Arterial Hypertension: Deep Sequencing Demonstrates Mitochondrial, Fibrotic, Inflammatory and Angiogenic Abnormalities.Applications:Unspecified applicationReactive species:Unspecified reactive speciesFrancois Potus et. al.PubMed 30213070
- Blood advances 2:1146-11562018TLR-activated repression of Fe-S cluster biogenesis drives a metabolic shift and alters histone and tubulin acetylation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWing-Hang Tong,Nunziata Maio,De-Liang Zhang,Erika M Palmieri,Hayden Ollivierre,Manik C Ghosh,Daniel W McVicar,Tracey A RouaultPubMed 29784770
Key facts
- Sample types
- Cell culture extracts, Tissue
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse, Rat, Cow, Human, Pig
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application FuncS | Reactivity Reacts | Dilution info - | Notes - |
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Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit is an immunoassay kit for the detection of Pyruvate dehydrogenase (PDH) Enzyme Activity in Mouse, Rat, Cow, Human, Pig in Cell culture extracts, Tissue samples.
Key facts
- Sample types
- Cell culture extracts, Tissue
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse, Rat, Cow, Human, Pig
- Assay Platform
- Reagents
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
ab109882 is used to quantify the activity of the PDH enzyme complex from human, bovine, mouse, rat and pig samples. This is accomplished by immunocapturing PDH with an anti-PDH antibody immobilized on a defined section of a dipstick. The enzyme complex is immunocaptured in its native form and activity is measured by the reaction scheme. PDH activity is visualized by coupling PDH-dependent production of NADH to the reduction of NBT in the presence of excess diaphorase, forming an insoluble intensely colored precipitate at the capture line. The signal intensity is measured by a dipstick reader or analyzed by other imaging systems such as a flatbed scanner.
Purified mitochondria can be used in this assay. However, homogenized tissue and whole cells can also be used without the need for mitochondrial isolation.
Pyruvate dehydrogenase activity is regulated by PDH kinase and PDH phosphatase. Cells grown in glucose media derive most of their energy from glycolysis; therefore, most of their PDH complex may be in an inactive phosphorylated form at the time of isolation. Growing cells in alternate carbon sources such as glutamine/galactose may up-regulate PDH activity. This kit does not include PDH kinases, PDH phosphatases or their respective inhibitors. These may be incorporated into the assay at the user's discretion.
Store dipsticks at room temperature out of direct sunlight in their provided desiccant container. Avoid high humidity conditions.
Store 20X Reagent Mix, Detergent, and Diaphorase at -80°C, but these may be aliquoted upon receipt to prevent freeze/thaw cycles.
Store NBT in the dark 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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Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit (ab109882)
Figure 2. An example using the ab109882 to measure PDH activity in human fibroblast sample.
Based on the standard curve, 160 µg of protein extract were loaded onto a dipstick for each sample. The figure shows two developed dipsticks, a control sample and a sample from a patient with a mutation in the E1-alpha subunit of PDH. The analysis of the signal intensity and interpolation from the standard curve showed that the patient sample had 8% the PDH activity of the control.
Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit (ab109882)
Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.
Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit (ab109882)
Figure 1. An example using the ab109882 to measure PDH activity in human fibroblast sample.
Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 160 µg of fibroblast protein extract in a 100 µL reaction volume (50 µL sample in Sample buffer plus 50 µL of Blocking solution). The dipsticks were developed for 1 hour in Activity buffer.
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