Antioxidant Assay Kit

Catalog No. ABIN1000278

Quick Overview for Antioxidant Assay Kit (ABIN1000278)

Target: Antioxidant

Application: Biochemical Assay (BCA)

Sample Type: Beverages, Food, Plasma, Saliva, Serum, Urine

Product Details

Specificity: 1.5 μM

Characteristics: Sensitive and accurate. Use 20 µL sample. Linear detection range from 1.5 to 1000 µM Trolox equivalents.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 10 min. Can be readily automated as a high-throughput assay for thousands of samples per day.

Components: Reagent A: 12 mL. Reagent B: 1 mL. Standard: 100 µL 50 mM Trolox.

Material not included: Pipetting devices, centrifuge tubes, clear flat-bottom uncoated 96-well plates, plate reader capable of reading optical density at 570nm, homogenizer or sonicator etc.

Application Details

Application Notes: Direct Assays: serum, plasma, urine, saliva and other biological samples, food and beverages.
Drug Discovery/Pharmacology: effects of drugs on TAC.

Protocol: 1. Standards and Samples. Equilibrate all components to room temperature. Briefly centrifuge Reagent B and Standard before opening. Mix 5 µL of the standard with 245 µL dH2O (final 1 mM Trolox). Dilute standards. Transfer 20 µL standards into wells of a clear flat-bottom 96-well plate. Transfer 20 µL of each sample into separate wells of the 96-well plate. Note: for unknown samples, perform several dilutions to ensure that TAC is within the linear range of1.5 to 1000 µMTrolox equivalents.
2. Assay. Prepare enough Working Reagent for Sample and Standard wells by mixing, for each assay well, 100 µL Reagent A and 8 µL Reagent B. Add 100 µL Working Reagent to all assay wells. Tap plate to mix. Incubate 10 min at room temperature.
3. Read OD570nm on a plate reader. Note: if calculated TAC is higher than 1000 µMTrolox equivalents, dilute sample in dH2O and repeat assay. Multiply the results by the dilution factor.

Restrictions: For Research Use only

Handling

Storage: -20 °C

References

Hadžović-Džuvo, Lepara, Valjevac, Avdagić, Hasić, Kiseljaković, Ibragić, Alajbegović: "Serum total antioxidant capacity in patients with multiple sclerosis." in: Bosnian journal of basic medical sciences / Udruženje basičnih mediciniskih znanosti = Association of Basic Medical Sciences, Vol. 11, Issue 1, pp. 33-6, (2011) (PubMed).

Habib, Eisa, Arafat, Marie: "Pulmonary involvement in early rheumatoid arthritis patients." in: Clinical rheumatology, Vol. 30, Issue 2, pp. 217-21, (2011) (PubMed).

Target Details

Target: Antioxidant

Background: Quantitative determination of total antioxidant capacity (TAC) by colorimetric (570nm) method.
Procedure: 20 min.

An Antioxidant is a molecule capable of slowing or preventing the oxidation of other molecules. Antioxidants protect the cells from damages by reactive oxygen species which are produced in oxidation reactions in the cell. Antioxidants can be small molecules such as glutathione, vitamins, or macromolecules such as catalase, glutathione peroxidase. As oxidative stress contributes to the development of many diseases including Alzheimer's disease, Parkinson's disease, diabetes, rheumatoid arthritis and neurodegeneration, the use of antioxidants in pharmacology is intensively studied. Antioxidants are also widely used as dietary supplements and in industry as preservatives in food, cosmetics, rubber and gasoline. Simple, direct and high-throughput assays for total antioxidant capacity (TAC) find wide applications in research, food industry and drug discovery. This improved assay measures total antioxidant capacity in which Cu 2+ is reduced by antioxidant to Cu + . The resulting Cu + specifically forms a colored complex with a dye reagent. The color intensity at 570nm is proportional to TAC in the sample.