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Product name
Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit
See all Pyruvate dehydrogenase products (6) ...
Tests
1 x 96 well plate
Sample type
Cell culture extracts, Tissue
Assay type
Enzyme activity
Cross reactivity
Reacts 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.
Notes
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.
Store 5X Stabilizer and 100X Reagent Dye at -20°C.
Store 20X Reagent Mix and 100X Coupler at -80°C.
Tested applications
Functional Studiesmore details
Properties
Storage instructions
Please see Notes section
| 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 |
Research areas
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Energy transfer pathways >> Energy Metabolism
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Carbohydrate metabolism
Metabolism >> Pathways and Processes >> Mitochondrial Metabolism >> Mitochondrial markers
Cancer >> Cancer Metabolism >> Metabolic signaling pathway >> Metabolism of carbohydrates
Kits/ Lysates/ Other >> Kits >> Cell Metabolism Kits >> Other Metabolism Assay
Signal Transduction >> Metabolism >> Mitochondrial
Signal Transduction >> Metabolism >> Energy Metabolism
Alternative names
- MSP18
Applications
Show applications key
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.
FuncS
Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit images:
Functional Studies - Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.
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, substrate is added, and enzyme activity is analyzed by measuring the change in absorbance of either the substrate or the product of the reaction (depending upon which enzyme is being analyzed). By analyzing the enzyme's activity in an isolated context, outside of the cell and free from any other variables, an accurate measurement of the enzyme's functional state can be understood.
- Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
Figure 1. PDH activity assay ab109902 reaction scheme.
- Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
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.
- Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
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.
Protocols
References for Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)
This product has been referenced in:
- Rapoport M, Salman L, Sabag O, Patel MS, Lorberbou Successful TAT-mediated enzyme replacement therapy in a mouse model of mitochondrial E3 deficiency. J Mol Med (Berl) 89:161-170 (2011).Read more (PubMed: ) »
- Yi X et al. Genetic reduction of lipoic acid synthase expression modestly increases atherosclerosis in male, but not in female, apolipoprotein E-deficient mice. Atherosclerosis 211:424-30 (2010).Read more (PubMed: 20347443) »
See all 7 publications for this product
Publishing research using ab109902? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"