{"product_id":"abcam-ab109902","title":"Abcam, ab109902, Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit","description":"\u003cp\u003eSize: 96Test\u003cbr\u003e\nPyruvate 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 \u0026gt; 100 publications\u003cbr\u003e\nKey facts\u003cbr\u003e\nDetection method:Colorimetric,\u003cbr\u003e\nSample types:Tissue Lysate, Cell culture extracts, Purified mitochondria,\u003cbr\u003e\nReacts with:Mouse, Rat, Cow, Human,\u003cbr\u003e\nAssay type:Enzyme activity,\u003cbr\u003e\nAssay time:3h 30m,\u003cbr\u003e\nAssay Platform:Microplate reader\u003c\/p\u003e\n\n\u003cp\u003eProduct details:\u003cbr\u003e\nThe 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.\u003cbr\u003e\nHow the assay works\u003cbr\u003e\nIn 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.\u003cbr\u003e\nThis 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.\u003cbr\u003e\nThe 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.\u003cbr\u003e\nPyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit protocol summary:\u003cbr\u003e\n- Add samples, standards and positive control to wells\u003cbr\u003e\n- Incubate for 3 hours at room temperature\u003cbr\u003e\n- Add 200 µL of Assay Solution to each well\u003cbr\u003e\n- Measure Optical Density (OD450 nm) in a kinetic mode at RT for 30 minutes\u003cbr\u003e\nHow other researchers are using\u003cbr\u003e\nPyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit has been used in a variety of sample type including:\u003cbr\u003e\nMouse cardiomyocytes\u003cbr\u003e\nMouse muscle tissue samples\u003cbr\u003e\nHuman iPSC-derived cardiomyocytes\u003cbr\u003e\nReferences:\u003cbr\u003e\n1-Mao Y et al. 2023\u003cbr\u003e\n2-Brenmoehl J et al. 2023\u003cbr\u003e\n3-Srivastava S et al. 2023\u003cbr\u003e\nRelated and recommended products\u003cbr\u003e\nSee an alternative kit to quantify Pyruvate dehydrogenase (PDH) Enzyme Activity:\u003cbr\u003e\n- Pyruvate dehydrogenase (PDH) Enzyme Activity Dipstick Assay Kit\u003cbr\u003e\nab109882\u003cbr\u003e\nStore 20X Buffer, Detergent, and Microplate at 4°C.\u003cbr\u003e\nStore 5X Stabilizer, 20X Reagent Mix, 100X Reagent Dye and 100X Coupler at -80°C.\u003c\/p\u003e\n\n\u003cp\u003eProperties and Storage Information:\u003cbr\u003e\nShipped at conditions-Dry Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols\u003c\/p\u003e\n\n\u003cp\u003eSupplementary Information:\u003cbr\u003e\nThis supplementary information is collated from multiple sources and compiled automatically.\u003cbr\u003e\nPyruvate 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).\u003cbr\u003e\nBiological function summary\u003cbr\u003e\nThe 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.\u003cbr\u003e\nPathways\u003cbr\u003e\nPyruvate 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.\u003cbr\u003e\nPyruvate 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.\u003c\/p\u003e","brand":"Abcam","offers":[{"title":"Default Title","offer_id":46843543683241,"sku":"ab109902","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/ar\/products\/abcam-ab109902","provider":"Iright","version":"1.0","type":"link"}