ATP Synthase (Complex V) Human Profiling ELISA Kit (ab124539)
- Product nameATP Synthase (Complex V) Human Profiling ELISA KitSee all Complex V kits ...
- Detection methodColorimetric
Intra-assay Sample n Mean SD CV% Overall 4 2.3%
- Tests1 x 96 test
- Sample typeTissue Extracts, Cell Lysate
- Assay typeSandwich (qualitative)
- Sensitivity12 µg/ml
- Range12 µg/ml - 1000 µg/ml
- Assay durationMultiple steps standard assay
- Species reactivityReacts with: Human
Does not react withMouse, Rat
- Product overview
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.
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at +4°C. Please refer to protocols.
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 TMB Development Solution 1 x 12ml
- Research Areas
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.
ATP Synthase (Complex V) Human Profiling ELISA Kit images
Example sample control curve of serially titrated HepG2 extract in the working range of the assay.
Human HepG2 cells were cultured in NARTI Zalcitabine (ddC) for 6 days to ensure a significant effect on mitochondrial DNA replication and mitochondrial protein translation, respectively. The drug reduced mitochondrial DNA levels and hence mitochondrial protein expression. As a consequence the assembly of Complexes I, III and IV were severely affected. Note that loss of the two small mitochondrial DNA encoded subunits of Complex V (ATP synthase) does not affect overall assembly. Interestingly an increase in Complex II was induced as a consequence of I, III, IV loss possibly to up regulate mitochondrial citric acid cycle function.
Human HepG2 cells were cultured in chloramphenicol for 6 days to ensure a significant effect on mitochondrial DNA replication and mitochondrial protein translation, respectively. The antibiotic inhibited mitochondrial protein translation and assembly of Complexes I and IV but had no significant effect on Complex II, III or V.
References for ATP Synthase (Complex V) Human Profiling ELISA Kit (ab124539)
This product has been referenced in:
- Sansanwal P et al. Insights into novel cellular injury mechanisms by gene expression profiling in nephropathic cystinosis. J Inherit Metab Dis 33:775-86 (2010). Read more (PubMed: 20865335) »