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Ubiquinol-cytochrome-c reductase (Complex III) is the third enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. Ubiquinol-cytochrome-c reductase is a large protein complex of approximately 250,000 MW made up of 11 different subunits. The enzyme forms a dimer in the mitochondrial inner membrane. The catalytic subunit cytochrome b is 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. Complex III catalyses the pumping of protons across the inner membrane and transfer of electrons from ubiquinol to cytochrome c by a mechanism known as the Q cycle in which ubiquinol (Q) the electron carrier exists as a reduced, partially reduced or oxidized form.
ab124537 Cyt C Reductase (Complex III) human profiling kit is an in vitro enzyme-linked immunosorbent assay (ELISA) for the comparison of Complex III levels or profile in cell and tissue lysates. The assay employs a human Complex III specific antibody coated onto microplate well plate strips.
Samples are pipetted into the wells and Complex III present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-Complex III 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 Complex III 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 Blocking Buffer||1 x 6ml|
|10X Complex III Detector Antibody||1 x 1ml|
|10X HRP Label||1 x 1ml|
|20X Buffer||1 x 20ml|
|Complex III Microplate||1 unit|
|Extraction Buffer||1 x 15ml|
|HRP Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab124537 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.|
Example control sample curve of serially titrated HepG2 extract in the working range of the assay.