Total Antioxidant Power Microplate Assay


The Total Antioxidant Power Microplate Assay kit is for the quantitative determination of the total anti-oxidative power in biological fluids by a microplate assay. The Total Antioxidant Power Microplate Assay is for research use only; not intended for use in diagnostic procedures.

SKU: TAP39-K01 Categories: , ,

Total Antioxidant Power Microplate Assay

The Total Antioxidant Power Microplate Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.125 mM
Dynamic Range: 0.125 – 2.0 mM
Incubation Time: 3 minutes
Sample Type: Biological Fluids
Sample Size: 15 µl

Product manufactured in the USA

Assay Background

Oxidative stress has been implicated in a number of diseases such as atherosclerosis, chronic inflammatory disease, chronic renal failure, and cancer.  It is a condition where an imbalance exists between the production of reactive oxidizing species and the body’s ability to neutralize these intermediates, resulting in cellular damage. The body has designed several physiological responses to oxidative stress including counterbalances such as enzymes and variously functionalized molecules (see examples below) that effectively neutralize these damaging species. These antioxidants can be either water or lipid soluble, and are localized transiently throughout various tissues, cells and cell types. 

Enzymes: Superoxide Dismutase, Catalase, Glutathione Peroxidase
Large Molecules:  Albumin, Ferritin, Ceruloplasmin
Small Molecules: Ascorbic Acid, α-Tocopherol, β-Carotene, Uric Acid

Given the multiplicity of antioxidant pathways, their centrality in the prevention of oxidant stress, and the influences of lifestyle and nutritional supplements on an individual’s antioxidant capacity, it is important to be able to quantitatively measure the total antioxidant capacity or antioxidant power with biological specimens.

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Additional Information

Assay Principle

The reduction potential of the sample or standard effectively converts Cu+2 to Cu+1, thus changing the ion’s absorption characteristics. This reduced form of copper will selectively form a stable 2:1 complex with the chromogenic reagent with an absorption maximum at ca. 450 nm12-16. A known concentration of Trolox is used to create a calibration curve, with the data being expressed as mM Trolox equivalents or in µM copper reducing equivalents.
A study was performed to evaluate the correlation between the concentration of principal antioxidants in human serum and the value obtained for total antioxidant power using this method with the results presented in Figure 1. Multivariate analysis of the obtained results yields a very high significance. The results obtained with the Total Antioxidant Power Assay for a series of serum samples were also compared to the resistance to oxidation of the serum lipids in these samples. The results of this study also show that these two parameters are highly correlated. The higher the total antioxidant power, the more protected the serum lipids are from oxidation.

  1. Dilute both Samples and Standards 1:40 in the provided Dilution Buffer (e.g. 15 µL serum + 585 µL Dilution Buffer).
  2. Place 200 µL of diluted Samples or Standards in each well. Reagent Blanks (BLK) should contain Dilution Buffer in place of Standard or Sample.
  3. Read the plate at 450 nm for a reference measurement.
  4. Add 50 µL of Copper Solution to each well and incubate for 3 minutes at room temperature.
  5. Add 50 µL of Stop Solution.
  6. Read the plate a second time at 450 nm.


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