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This rapid multiplexing microplate kit is used to determine both the activity and quantity of cytochrome c oxidase in a human sample with greater speed and simplicity. The ratio of the two is a measure of specific activity. The COX enzyme is immunocaptured within the wells of the microplate and the activity is determined by following the oxidation of reduced Cytochrome c colorimetrically by the absorbance change at 550 nm. Subsequently, in these same wells the quantity of the enzyme can then be determined by adding a COX specific detector antibody and the appropriate alkaline phophatase conjugated antibody; this phosphatase changes a substrate from colorless to yellow at 405 nm. This reaction takes place in a time dependent manner proportional to the amount of protein captured in the wells. Included in this kit for performance of the multiplexing assay are buffers, detergent, substrates, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.
|20X Buffer||1 x 15ml|
|96-well microplate (12 strips)||1 unit|
|AP Development Reagent||1 x 400µl|
|AP label 2500X||Tube B||1 x 12µl|
|Detector antibody||Tube A||1 x 1ml|
|Detergent||1 x 1ml|
|Development buffer||Tube 3||1 x 10ml|
|Reagent C (Cytochrome c)||1 x 1ml|
|Wash Buffer||Tube 2||1 x 2ml|
Our Abpromise guarantee covers the use of ab109910 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.|