Description
Antioxidant Assay Kit (BA0059) (BA0059)
The Antioxidant Assay Kit (SKU: BA0059) provides a simple, direct and high-throughput means of measuring total antioxidant capacity (TAC) in a broad range of samples. Antioxidants are molecules capable of slowing or preventing the oxidation of other molecules, protecting cells from damage caused by reactive oxygen species. Because oxidative stress contributes to the development of many diseases, including Alzheimer's disease, Parkinson's disease, diabetes and rheumatoid arthritis, the use of antioxidants is intensively studied in pharmacology. This improved assay measures total antioxidant capacity in which copper(II) is reduced by antioxidant to copper(I), and the resulting copper(I) forms a coloured complex with a dye reagent. The colour intensity at 570 nm is proportional to the TAC in the sample.
| Product Name: | Antioxidant Assay Kit (BA0059) |
| SKU: | BA0059 |
| Detection Method: | Colorimetric |
| Detection Range: | 1.5 to 1000 uM Trolox equivalents |
| Sample Type: | Serum, plasma, urine, saliva and other biological samples, food and beverages |
| Species Reactivity: | All |
| Assay Time: | 10 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Room Temperature |
Quantitative colorimetric determination of total antioxidant capacity via reduction of copper(II) to copper(I), which forms a coloured complex with a dye reagent read at 570 nm.
- Sensitive and accurate; uses only 20 uL sample
- Linear detection range from 1.5 to 1000 uM Trolox equivalents
- Simple, high-throughput procedure involving a single working reagent and a 10 minute incubation
- Can be readily automated for thousands of samples per day
- Direct assays of serum, plasma, urine, saliva and other biological samples, food and beverages
- Drug discovery and pharmacology: effects of drugs on total antioxidant capacity
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Standards and Samples. Equilibrate all components to room temperature. Briefly centrifuge Reagent B and Standard before opening. Mix 5 uL standard with 245 uL distilled water (final 1 mM Trolox) and prepare the dilution series shown in the table. Transfer 20 uL standards into wells of a clear flat-bottom 96-well plate. Transfer 20 uL of each sample into separate wells; for unknown samples perform several dilutions to ensure the result falls within the linear range. |
| 2 | Assay. Prepare enough working reagent by mixing, for each well, 100 uL Reagent A with 8 uL Reagent B. Add 100 uL working reagent to all wells and tap to mix. Incubate for 10 minutes at room temperature. |
| 3 | Read the optical density at 570 nm on a plate reader. If the calculated TAC exceeds 1000 uM Trolox equivalents, dilute the sample in distilled water, repeat the assay and multiply by the dilution factor. |
Subtract the blank OD value (standard #4) from all standard and sample OD values. Plot the change in OD at 570 nm against standard concentrations and determine the slope. Calculate TAC = (ODsample - ODblank) / slope x n, where n is the sample dilution factor, expressed in uM Trolox equivalents.
| Component | Quantity | Storage |
| Reagent A | 12 mL | -20C |
| Reagent B | 1 mL | -20C |
| Standard (50 mM Trolox) | 100 uL | -20C |
