Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit is an immunoassay kit for the detection of Pyruvate dehydrogenase (PDH) Protein Quantity in Mouse, Rat, Cow, Human in Cell culture extracts, Tissue samples.
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Key facts
- Sample types
- Cell culture extracts, Tissue
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse, Rat, Cow, Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit is an immunoassay kit for the detection of Pyruvate dehydrogenase (PDH) Protein Quantity in Mouse, Rat, Cow, Human in Cell culture extracts, Tissue samples.
Key facts
- Sample types
- Cell culture extracts, Tissue
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse, Rat, Cow, Human
- Assay Platform
- Reagents
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
ab109883 (MSP31) is used to quantify the activity of the PDH enzyme complex (pyruvate dehydrogenase) from human, bovine, mouse, and rat samples.
PDH is present in all tissues; it plays a central role in metabolism as it is a key regulatory enzyme that functions at the junction between glycolysis and the tricarboxylic acid cycle. The PDH complex is composed of multiple copies of three catalytic component enzymes; pyruvate dehydrogenase or E1 (EC 1.2.4.1), dihydrolipoamide transacetylase or E2 (EC 2.3.1.12) and dihydrolipoamide dehydrogenase or E3 (EC 1.8.1.4). Inherent to its regulatory function, a number of other proteins regulate PDH activity. One of these, dihydrolipoamide dehydrogenase-binding protein (E3Bp) is necessary for the interaction of the E2 and E3 components.
This dipstick assay kit utilizes two monoclonal antibodies. One recognizes the E2 enzyme (monoclonal antibody bound to the dipstick) and the other antibody (gold-conjugated) recognizes both the E2 and E3Bp components of the PDH complex. The signal intensity is measured by a dipstick reader or analyzed by other imaging systems such as a flatbed scanner. The kit is compatible with a variety of sample types (tissue or cell culture) from a number of different species (human, bovine, mouse, and rat).
All components are stable in their provided containers at room temperature out of direct sunlight.
After diluting the 10X Blocking Buffer to 2X, store at 4°C.
For long-term storage, all buffers can be stored at 4°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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3 product images
ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
An example using ab109883 to quantify PDH levels using various concentration of rat tissue protein extract.
Based on the above standard curve values, 18 µg (control - undiluted), 9 µg (sample A - diluted 1:2) and 1.8 µg (sample B - diluted 1:10) of extract were loaded to each dipstick. Two dipsticks per sample were run (typical intra-assay precision is 90-98%). Using GraphPad software, the signal intensity from the standard curve was interpolated and the quantity of PDH in samples A and B was determined. Based on the above analysis, the unknown samples, A and B, had 48% and 9% of control PDH levels, respectively.
ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
An example using ab109883 to quantify PDH levels using various concentration of rat tissue protein extract.
Shown is a 1:2 dilution series using a positive control sample. Approximately 7 to 8 dipsticks are suitable for covering the entire working range and the blank for background levels. In this example the dilution series starts with 20 µg of rat heart tissue extract. A one-site hyperbola line was generated for best-fit analysis using GraphPad.
Sandwich ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted, and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. Multiplexing dipstick assays have multiple target protein lines. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurred.
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