Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
- Product nameComplex IV Human Enzyme Activity Dipstick Assay KitSee all Complex IV kits ...
- Sample typeCell culture extracts, Tissue
- Assay typeQuantitative
- Species reactivityReacts with: Cow, Human
- Product overview
Contains 30 or 90 dipsticks and necessary components to quantify the activity of the cytochrome c oxidase enzyme complex from human and bovine samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.
The isolation of mitochondria is not necessary for the performance of this assay. In this kit the specificity of anti-COX monoclonal antibodies (mAbs) is combined with traditional methods for determining COX enzyme activity by histochemical methods and in-gel activity assays. First, the COX enzyme complex is immnocaptured (i.e immunoprecipated in active form) on the dipstick. Second, the dipstick is immersed in COX activity buffer containing reduced cytochrome c and di-amino benzidinetetrachloride (DAB), which serves as the reporter of COX activity. Immunocaptured COX oxidizes cytochrome c, which then oxidizes DAB to form a red-colored precipitate at the COX antibody line on the dipstick. In addition to being quick, the reaction is cyanide-sensitive. The signal intensity of this precipitate corresponds to the level of COX activity in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.
Store dipsticks at room temperature in their provided container and out of direct sunlight. High humidity conditions should be avoided.
Store Buffer A, B, and C at 4°C or at -20°C for long term storage.
Store Tubes 1 and 2 at -80°C; they can also be aliquoted upon receipt to prevent freeze/thaw cycles.
Tube 3 can be stored at room temperature.
- Tested applicationsFunctional Studies more details
- Storage instructionsPlease refer to protocols.
Components 90 tests 30 tests 96-well microplate 6 units 2 units Buffer B (Blocking solution) 1 x 6ml 1 x 2ml Buffer C (Wash buffer) 1 x 3ml 1 x 1ml Dipsticks 1 x 90 units 1 x 30 units Extraction Buffer 1 x 45ml 1 x 15ml Tube 1 (DAB 100X stock) 1 x 300µl 1 x 100µl Tube 2 (reduced cytochrome c 20X stock) 1 x 1.5ml 1 x 500µl Tube 3 (Buffer for activity assay) 1 x 30ml 1 x 10ml
- Research Areas
- COXCytochrome c oxidaseCytochrome oxidase
- Entrez Gene: 4512 Human
- Entrez Gene: 4513 Human
- Entrez Gene: 4514 Human
- Entrez Gene: 1345 Human
- Entrez Gene: 1351 Human
- Entrez Gene: 1329 Human
- Entrez Gene: 1349 Human
- Entrez Gene: 1337 Human
- Entrez Gene: 1327 Human
- Entrez Gene: 1340 Human
- Entrez Gene: 1350 Human
- Entrez Gene: 9377 Human
- Entrez Gene: 1346 Human
- SwissProt: P00395 Human
- SwissProt: P00403 Human
- SwissProt: P00414 Human
- SwissProt: P13073 Human
- SwissProt: P12074 Human
- SwissProt: P15954 Human
- SwissProt: P24311 Human
- SwissProt: P10176 Human
- SwissProt: P14854 Human
- SwissProt: P09669 Human
- SwissProt: P24310 Human
- SwissProt: P20674 Human
- SwissProt: P10606 Human
Our Abpromise guarantee covers the use of ab109876 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Complex IV Human Enzyme Activity Dipstick Assay Kit images
Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.
Figure 1. An example using ab109876 to measure Complex IV activity in fibroblast protein extracts. Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 100 µg of fibroblast protein extract.
Figure 2. An example using ab109876 to measure Complex IV activity in fibroblast protein extracts. Based on the standard curve, 50 µg of protein extract were loaded onto a dipstick for each sample. The figure shows four developed dipsticks, a control sample (1) and four unknowns (2-6). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 15-61% of normal Complex IV activity levels.
References for Complex IV Human Enzyme Activity Dipstick Assay Kit (ab109876)
This product has been referenced in:
- Calvo SE et al. Molecular diagnosis of infantile mitochondrial disease with targeted next-generation sequencing. Sci Transl Med 4:118ra10 (2012). Read more (PubMed: 22277967) »
- Fassone E et al. FOXRED1, encoding an FAD-dependent oxidoreductase complex-I-specific molecular chaperone, is mutated in infantile-onset mitochondrial encephalopathy. Hum Mol Genet 19:4837-47 (2010). Read more (PubMed: 20858599) »