QuantiChrom™ ABTS Antioxidant Assay Kit

Protocol SDS

Application

• For quantitative determination of antioxidant capacity in food, beverage, and urine samples.

Key Features

• Fast and convenient. 5-minute reagent incubation period allows for quick sample testing.
• Adaptable. Compatible with aqueous and ethanol extracted samples, and stable signal allows for extended run-time length (up to 2 hours).
• High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated to assay thousands of samples per day.

Method

• OD650nm

Samples

• Food, beverage, and urine samples

Species

• All

Procedures

• 35 min

Size

• 100 tests in 96-well plate

Detection Limit

• 20 μM Trolox equivalents

Shelf Life

• 6 months

More Details

2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS^•+) is a green cation radical that returns to its colorless state when reduced. The ABTS assay is widely used to measure antioxidant capacity, which is the ability of redox molecules in foods and biological systems to scavenge free radicals. BioAssay Systems’ ABTS antioxidant assay measures the change in absorbance that occurs when the ABTS radical is reduced and decolorized from green to clear. The decrease in absorbance at 650 nm is directly proportional to the antioxidant concentration and is compared to a Trolox antioxidant standard curve.

Do I need to include a sample blank in my testing?

Most samples will not have enough background OD at 650 nm to require a sample blank. Typically samples with color are diluted to the point where the background is insignificant. For example, in our red wine testing we diluted the wine 100x and the background was insignificant. In the case that a sample background is needed, prepare a blank working reagent (BWR) by combining in a 1.5 mL tube, for each sample blank well, 190 μL ABTS Reagent + 1 μL HRP. Incubate 5 minutes at room temperature along with WR, and add 180 μL BWR to 20 μL sample. For the calculation use the formula: [Trolox] = (OD_WATER – OD_SAMPLE – OD_SAMPLEBLANK) / slope x n (μM)

Can I read the assay at 420 nm?

Our assay has been optimized to be read at 650 nm to avoid background OD, but the assay can be read at 420 nm. In this case all assay volumes must be cut in half due to the higher signal at 420 nm (add 90 μL WR to 10 μL sample).

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.