Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit

Catalog No. MSP18

$425.00 - 96 tests


Product Description

This rapid activity microplate kit is used to determine the activity of PDH in a human, bovine, mouse, or rat sample. 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. Therefore, for studies with cultured cells, MitoSciences suggests using the MSP30 Dipstick Assay Kit for PDH Activity, which has a working range of 100-1000 µg depending on the cell type.



 
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Product Specifications
 
Species Reactivity: human, rat, mouse, bovine
Storage Conditions: Store 20X Reagent Mix and 100X Coupler at -80°C.

Store 5X Stabilizer and 100X Reagent Dye at -20°C.

Store all other components at 4°C.
Country of Origin: USA


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Figure 1


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Figure 1. PDH activity assay MSP18 reaction scheme.

Figure 2


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Figure 2. Example of microplate reader recorded data. Bovine heart mitochondria were loaded at 100 µg/well (top trace) and 2-fold dilutions (stepwise lower traces). Activity should always be related to a control or normal sample to obtain the relative activity of PDH in experimental samples.

Figure 3


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Figure 3. 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.


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Downloadable Documents


   Technical Data Sheet

   Protocol

   PDH Playbook


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Published Studies Using This Product: Lee et al., 2009. Inhibition of prolyl hydroxylase protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity: model for the potential involvement of the hypoxia-inducible factor pathway in Parkinson disease.

Sikder et al., 2007. The neurohormone orexin stimulates hypoxia-inducible factor-1 activity.


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