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Total Antioxidant Capacity Assay Kit (ab65329) can measure either the combination of both small molecule antioxidants and proteins or small molecules alone in the presence of our proprietary Protein Mask. Cu2+ ion is converted to Cu+ by both small molecule and protein. The Protein Mask prevents Cu2+ reduction by protein, enabling the analysis of only the small molecule antioxidants. The reduced Cu+ ion is chelated with a colorimetric probe giving a broad absorbance peak around 570 nm, proportional to the total antioxidant capacity.
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Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
Antioxidants play an important role in preventing the formation of and scavenging of free radicals and other potentially toxic oxidizing species. There are three categories of antioxidant species: enzyme systems (GSH reductase, catalase, peroxidase, etc.), small molecules (ascorbate, uric acid, GSH, vitamin E, etc.) and proteins (albumin, transferrin, etc.). Different antioxidants vary in their reducing power. Trolox is used to standardize antioxidants, with all other antioxidants being measured in Trolox equivalents. Measurement of the combined non-enzymatic antioxidant capacity of biological fluids and other samples provides an indication of the overall capability to counteract reactive oxygen species (ROS), resist oxidative damage and combat oxidative stress-related diseases. In some cases, the antioxidant contribution of proteins is desired whereas in other cases only the contribution of the small molecule antioxidants is needed.
|Assay Diluent||WM||1 x 10ml|
|Cu++ Reagent||Blue||1 x 0.2ml|
|Protein Mask||NM||1 x 10ml|
|Trolox Standard (1 µmole) (Lyophilized)||Yellow||1 vial|
Our Abpromise guarantee covers the use of ab65329 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.|
Total antioxidant capacity was determined in mouse cardiac homogenates using Total antioxidant capacity assay kit (ab65329).
Trolox equivalent capacity measured in milk and concentrated squash. Background signal subtracted, duplicates; +/- SD
Trolox equivalent capacity measured in mouse tissue lysates, showing quantity (nmol) per mg of extracted protein. Results following blocking of protein activity is shown (Mask). (Duplicates; +/- SD).
Trolox equivalent capacity measured in biological fluids. Results following blocking of protein activity is shown (Mask). Background signal subtracted, duplicates; +/- SD.