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Pyruvate 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. 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.
Store 20X Buffer, Detergent, and Microplate at 4°C.
Store 5X Stabilizer at -20°C.
Store 20X Reagent Mix, 100X Reagent Dye and 100X Coupler at -80°C.
|20X Buffer||1 x 15ml|
|20X Reagent Mix||2 x 600µl|
|5X Stabilizer||1 x 13ml|
|96-well Microplate (12 strips)||1 unit|
|Coupler||1 x 250µl|
|Detergent||2 x 1ml|
|Reagent Dye||1 x 250µl|
Our Abpromise guarantee covers the use of ab109902 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|