Abcam’s Succinate dehydrogenase (Complex II) Profiling in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for comparison of Succinate dehydrogenase levels in Human cell and tissue lysates.
The assay employs a Human succinate dehydrogenase complex specific capture coated onto microplate well plate strips. Samples are pipetted into the wells and succinate dehydrogenase present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-succinate dehydrogenase 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 succinate dehydrogenase (Complex II) 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.
Succinate dehydrogenase (Complex II) is the second enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. Succinate dehydrogenase is a large protein complex of 140,000 MW made up of 4 different subunits (SDH, SDB, SDHC, and SDHD). The enzyme forms a dimer in the mitochondrial inner membrane. Succinate dehydrogenase is the only enzyme of the OXPHOS system containing no mitochondrial DNA (mtDNA) encoded subunit components. Therefore all four subunits are encoded by nuclear genomic DNA, made in the cytosol, and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from succinate via a flavin adenine dinucleotide (FAD), three iron sulfur clusters and b-type cytochrome. Succinate dehydrogenase also participates in the citric acid cycle producing fumarate for the next step in the cycle - hydration by fumarase.
|Components||1 x 96 tests|
|10X Blocking Buffer||1 x 6ml|
|10X HRP Label||1 x 1ml|
|10X SDH Detector Antibody||1 x 0.75ml|
|20X Buffer||1 x 20ml|
|Extraction Buffer||1 x 15ml|
|SDH Microplate||1 unit|
|HRP Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab124536 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.|
ab124536 has not yet been referenced specifically in any publications.