Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
Product nameTotal Antioxidant Capacity Assay Kit
Sample typeUrine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
Assay time2h 00m
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.
In the total antioxidant capacity assay protocol, the Cu2+ ion is converted to Cu+ by both small molecule and protein antioxidants. The Protein Mask prevents Cu2+ reduction by proteins, 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.
Total antioxidant capacity assay protocol summary:
- add protein mask to samples if only measuring small molecule total antioxidant capacity
- add samples and standards to wells
- add Cu2+ solution and incubate for 90 min at room temp
- analyze with microplate reader
Antioxidants play an important role in preventing the formation of, and scavenging of, free radicals and other 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.).
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.
Review the oxidative stress marker and assay guide to learn about more assays for oxidative stress.
Storage instructionsStore at +4°C. Please refer to protocols.
Components Identifier 100 tests Assay Diluent WM 1 x 10ml Cu++ Reagent Blue 1 x 0.2ml Protein Mask NM 1 x 10ml Trolox Standard (1 µmol) (Lyophilized) Yellow 1 vial
RelevanceAntioxidants 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, and cooperation of all different antioxidants provides greater protection against reactive oxigen or nitrogen radicals than any single compound alone. Therefore, the overall total antioxidant capacity may give more relevant biological information compared to that obtained by the measurement of individual components, as it considers the cumulative effect of all antioxidants present.
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.
ab65329 has been referenced in 50 publications.
- Madariaga VI et al. Myogenous temporomandibular disorders and salivary markers of oxidative stress-A cross-sectional study. J Oral Rehabil 48:1-9 (2021). PubMed: 32979853
- Li M et al. Brachyury engineers cardiac repair competent stem cells. Stem Cells Transl Med 10:385-397 (2021). PubMed: 33098750
- Okda TM et al. Chemopreventive and anticancer activities of indomethacin and vitamin D combination on colorectal cancer induced by 1,2-dimethylhydrazine in rats. Biomed Rep 14:27 (2021). PubMed: 33408861
- Shili CN et al. Effect of a Phytogenic Water Additive on Growth Performance, Blood Metabolites and Gene Expression of Amino Acid Transporters in Nursery Pigs Fed with Low-Protein/High-Carbohydrate Diets. Animals (Basel) 11:N/A (2021). PubMed: 33672517
- Gupta SK et al. Effect of rutin on retinal VEGF, TNF-a, aldose reductase, and total antioxidant capacity in diabetic rats: molecular mechanism and ocular pharmacokinetics. Int Ophthalmol 40:159-168 (2020). PubMed: 31456155