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ab118184, is used to determine the relative specific activity (activity and quantity) of catalase in a sample. The native enzyme is immunocaptured within the wells of the microplate; this removes all other enzymes. The assay buffer contains hydrogen peroxide which reacts with a substrate to make a luminescent product. Catalase functions rapidly to remove hydrogen peroxide from the solution and reduce the production of light. Therefore the light produced in each well is inversely proportional to the amount of catalase activity.
After activity measurement the quantity of catalase is measured by adding to each well an anti-catalase detector antibody. After washing away unbound detector antibody, HRP-conjugated label specific for the detector antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of catalase 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.
|Components||1 x 96 tests|
|10X Blocking Solution||1 x 10ml|
|10X Detector Antibody||1 x 1.5ml|
|10X HRP Label||1 x 1.5ml|
|200X Luiminescent reagent||1 x 0.2ml|
|20X Buffer||1 x 20ml|
|55X Coupler||1 x 0.5ml|
|60X Hydrogen Peroxide||1 x 0.5ml|
|96-Well Microplate||1 unit|
|Base Buffer||1 x 24ml|
|Extraction Buffer||1 x 15ml|
|HRP Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab118184 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 concentration.|