Pyruvate dehydrogenase (PDH) Profiling ELISA Kit
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(3 Publications)
- ELISA
Supplier Data
ELISA - Pyruvate dehydrogenase (PDH) Profiling ELISA Kit (AB110174)
An example of the quantity of PDH capture from a HepG2 cultured cell lysate. The sample was diluted to show that over this range of concentrations that can be used. Each sample was measured in 6 replicates. Bars show standard deviations.
- sELISA
Supplier Data
Sandwich ELISA - Pyruvate dehydrogenase (PDH) Profiling ELISA Kit (AB110174)
Abcams' protein quantity microplate assays use the well-established sandwich ELISA format, whereby capture and detector antibodies are used to immobilize and then quantify a target protein or enzyme. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state. Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, a second monoclonal antibody, against a different epitope on the target, is added to the well. This detector antibody is either directly labeled with biotin, or a biotin-labeled goat anti-mouse secondary is added. Substrate plus HRP or AP conjugated to streptavidin provide a colorimetric signal that is readable by any plate readers capable of standard ELISA absorbance measurements.
Reactivity data
Product details
Abcam's Pyruvate dehydrogenase (PDH) in vitro ProfilingELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the measurement of Pyruvate dehydrogenase (PDH) in Human, bovine, mouse, and rat whole tissue or cell lysate samples.
Capture antibodies are pre-coated in the wells of modular microplates, which can be broken into 8-well strips. This assay is a "sandwich" ELISA, where the PDH enzyme is purified and immobilized by an anti-PDH capture antibody pre-coated in the microplate wells. The amount of captured PDH is determined by adding a second (detector) anti-PDH antibody which binds to the captured PDH hat a different epitope. This is followed by binding of an HRP conjugated goat anti-mouse antibody that binds the detector anti-PDH antibody. The detector-bound HRP then changes the colorless HRP development solution to blue and the color intensity (absorbance) is proportional to the amount of PDH captured. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state.
5X Stabilizer should be stored at -20°C. Remainder of kit should be stored at 4°C. When stored as recommended the kit is stable for 6 months.
PDH is the key regulatory enzyme of cellular metabolism because it links the TCA cycle and subsequent oxidative phosphorylation with glycolysis and gluconeogenesis as well as with both lipid and amino acid metabolism. PDH activity is regulated primarily by PDK-dependent phosphorylation and PDP-dependent dephosphorylation of PDH. Phosphorylation inactivates PDH whereas dephosphorylation activates PDH. Phosphorylation occurs at Serines 232, 293, and 300 of the human E1α subunits.
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Supplementary information
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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.
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Publications (3)
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Cancer research 83:3478-3491 PubMed37526524
2023
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Unspecified application
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Journal of the American Society of Nephrology : JASN 25:998-1012 PubMed24385590
2014
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ELISA
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Mouse
The Journal of clinical investigation 122:267-79 PubMed22201682
2011
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