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ATP synthase (Complex V) is the fifth enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. ATP synthase is a large protein complex of approximately 550,000 MW made up of 17 different subunits arranged in a membrane embedded proton translocating domain (F0 domain) and a soluble ATP synthesizing catalytic domain (F1 domain). Two membrane embedded subunits ATP6 (subunit a) and ATP8 (subunit 8) are encoded on mitochondrial DNA (mtDNA). All other subunits are encoded by nuclear genomic DNA, made in the cytosol, and translocated into the organelle for assembly at the inner membrane. A proton gradient generated by the other enzymes of the OXPHOS system provides the energy for ATP synthesis by a rotational catalytic method known as the binding change mechanism. ATP synthase is regulated by CabI and IF1 proteins in response to calcium concentration and mitochondrial membrane potential respectively.
ab124539 ATP synthase (Complex V) human profiling kit is an in vitro enzyme-linked immunosorbent assay (ELISA) for the comparison of ATP synthase levels or profile in cell and tissue lysates. The assay employs a capture antibody specific for human ATP synthase coated onto microplate well strips.
Samples are pipetted into the wells and ATP synthase present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-ATP synthase detector antibody is added. After washing away unbound detector antibody, an HRP-conjugated secondary antibody specific for the detector antibody is pipetted into the wells. The wells are again washed, an HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of ATP synthase bound. The developing blue color is measured at 600 nm.
Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Species– Human. Rat and Mouse samples are not appropriate, other species are untested.
Store all components at 4°C. This kit is stable for 6 months from receipt. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
|Components||1 x 96 tests|
|10X ATP Synthase Detector Antibody||1 x 1ml|
|10X Blocking Buffer||1 x 6ml|
|10X HRP Label||1 x 1ml|
|20X Buffer||1 x 20ml|
|ATP Synthase Microplate||1 unit|
|Extraction Buffer||1 x 15ml|
|HRP Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab124539 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 concentration.|