Application

Antioxidant assay kit provides all of the reagents required for an efficient measurement of the total antioxidant capacity of plasma, serum, urine, saliva, cells, and tissue lysates. It has also been used to measure the radical cation (2,2′-azino-di-[3-ethylbenzthiazoline sulphonate]) (ABTS+) scavenging capacity.

It was tested on A431 and CHO cell lysates; rat brain, liver, and kidney lysates; human plasma, serum, urine, and saliva.

General description

Free radicals or reactive oxygen species (ROS) are produced during biochemical redox reactions as part of normal physiological cell metabolism (protection from infectious organisms) and as a response to environmental factors such as UV light, cigarette smoke, environmental pollutants, and g-radiation. Once formed, ROS attack cellular components causing damage to lipids, proteins, and DNA, which can initiate numerous diseases, including cancer, atherosclerosis, rheumatoid arthritis, diabetes, liver damage, and central nervous system disorders.

In the presence of antioxidants the radical cation is suppressed to an extent dependent on the activity of the antioxidant and the color intensity is decreased proportionally. Trolox, a water-soluble vitamin E analogue, serves as a standard or a control antioxidant.

Preparation Note

Reagents are to be prepared with ultrapure water.

Principle

The principle of the antioxidant assay is formation of a ferryl myoglobin radical from metmyoglobin and hydrogen peroxide, which oxidizes the ABTS (2,2¢-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) to produce a radical cation, ABTS·+ , a soluble chromogen that is green in color and can be determined spectrophotometrically at 405 nm.

Reconstitution

Myoglobin Solutions are to be stored at -20C after reconstitution.Trolox Working Solution is to be stored at -20C after reconstitution.ABTS Substrate Solution is to be stored at 4C for 2 weeks or at -20C for 1 month.

Suitability

Suitable for the measurement of the total antioxidant capacity of plasma, serum, urine, saliva, cells, and tissue lysates