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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.
|组件||90 tests||30 tests|
|10X Blocking Buffer||6 x 1ml||2 x 1ml|
|96-well microplate||6 units||2 units|
|Dipsticks||1 x 90 units||1 x 30 units|
|Extraction Buffer||3 x 15ml||1 x 15ml|
|Tube 1 (DAB 100X stock)||3 x 100µl||1 x 100µl|
|Tube 2 (reduced cytochrome c 20X stock)||3 x 500µl||1 x 500µl|
|Tube 3 (Buffer for activity assay)||3 x 10ml||1 x 10ml|
|Wash buffer||3 x 1.5ml||1 x 1.5ml|
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.
|Functional Studies||Use at an assay dependent dilution.|