Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
- Product namePyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay KitSee all Pyruvate dehydrogenase kits ...
- Detection methodColorimetric
- Tests1 x 96 well plate
- Sample typeCell culture extracts, Tissue
- Assay typeEnzyme activity
- Assay durationMultiple steps standard assay
- Species reactivityReacts with: Mouse, Rat, Cow, Human
- Product overview
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.
Store 20X Buffer, Detergent, and Microplate at 4°C.
Store 5X Stabilizer and 100X Reagent Dye at -20°C.
Store 20X Reagent Mix and 100X Coupler at -80°C.
- Tested applicationsFunctional Studies more details
- Storage instructionsPlease refer to protocols.
Components 96 tests 100X Coupler 1 x 250µl 100X Reagent Dye 1 x 250µl 20X Buffer 1 x 15ml 20X Reagent Mix 2 x 600µl 5X Stabilizer 1 x 13ml 96-well Microplate (12 strips) 1 unit Detergent 2 x 1ml
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.
Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit images
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.
Example of microplate reader recorded data from bovine heart mitochondria (100 µg/well) (top trace) and 2-fold dilutions (stepwise lower traces) using Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
Schematic diagram showing the reaction involved in Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
Principle of Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
References for Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
This product has been referenced in:
- Morten KJ et al. Differential regulation of HIF-mediated pathways increases mitochondrial metabolism and ATP production in hypoxic osteoclasts. J Pathol 229:755-64 (2013). Read more (PubMed: 23303559) »
- Phillips BE et al. Effect of colon cancer and surgical resection on skeletal muscle mitochondrial enzyme activity in colon cancer patients: a pilot study. J Cachexia Sarcopenia Muscle 4:71-7 (2013). Functional Studies ; Human . Read more (PubMed: 22648738) »