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Abcam’s Pyruvate dehydrogenase (PDH) in vitro ProfilingELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the measurement of Pyruvate dehydrogenase (PDH) in Human, bovine, mouse, and rat whole tissue or cell lysate samples.
Capture antibodies are pre-coated in the wells of modular microplates, which can be broken into 8-well strips. This assay is a “sandwich” ELISA, where the PDH enzyme is purified and immobilized by an anti-PDH capture antibody pre-coated in the microplate wells. The amount of captured PDH is determined by adding a second (detector) anti-PDH antibody which binds to the captured PDH hat a different epitope. This is followed by binding of an HRP conjugated goat anti-mouse antibody that binds the detector anti-PDH antibody. The detector-bound HRP then changes the colorless HRP development solution to blue and the color intensity (absorbance) is proportional to the amount of PDH captured. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state.
5X Stabilizer should be stored at -20°C. Remainder of kit should be stored at 4°C. When stored as recommended the kit is stable for 6 months.
PDH is the key regulatory enzyme of cellular metabolism because it links the TCA cycle and subsequent oxidative phosphorylation with glycolysis and gluconeogenesis as well as with both lipid and amino acid metabolism. PDH activity is regulated primarily by PDK-dependent phosphorylation and PDP-dependent dephosphorylation of PDH. Phosphorylation inactivates PDH whereas dephosphorylation activates PDH. Phosphorylation occurs at Serines 232, 293, and 300 of the human E1α subunits.
|10X Blocking Buffer||1 x 10ml|
|20X Buffer||1 x 20ml|
|20X Detector Antibody||1 x 1ml|
|20X HRP Label||1 x 1ml|
|5X Stabilizer||1 x 13ml|
|96-well Microplate (12 strips)||1 unit|
|Detergent||1 x 1ml|
|HRP/TMB Development Solution||1 x 20ml|
Our Abpromise guarantee covers the use of ab110174 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent dilution.|