Description

FRAP-Based Antioxidant Detection

The kit uses the FRAP (Ferric Reducing Antioxidant Power) method, where antioxidants reduce ferric ions (Fe³⁺) to ferrous ions (Fe²⁺), forming a blue Fe²⁺-TPTZ complex measurable at 593 nm.

Rapid Colorimetric Measurement

The reaction produces a blue chromogenic product that can be measured quickly using a microplate reader or spectrophotometer, providing reliable quantitative results.

Broad Sample Compatibility

The kit is designed to analyze antioxidant capacity in multiple biological matrices including:

• Serum
• Plasma
• Tissue homogenates
• Cell lysates or cell culture supernatants
• Saliva
• Urine

This versatility makes it suitable for a wide range of experimental designs.

Assay Principle

Under acidic conditions, antioxidants in the sample reduce Fe³⁺-TPTZ to Fe²⁺-TPTZ, producing a blue-colored complex.

The intensity of the blue color is proportional to the total antioxidant capacity of the sample and is measured spectrophotometrically at 593 nm.

The antioxidant capacity is then calculated by comparing the absorbance of the sample to a standard curve generated using FeSO₄ standard solutions.

Biological Significance

Cells and biological systems contain various antioxidant molecules and enzymes that neutralize reactive oxygen species (ROS) and protect against oxidative stress.

Measuring total antioxidant capacity provides valuable insight into:

• Oxidative stress levels
• Cellular defense mechanisms
• Metabolic and physiological conditions
• Antioxidant responses in biological systems

Therefore, evaluating T-AOC is an important tool in biochemical and biomedical research.

Kit Components

The kit includes the following reagents:

• Check Buffer (R1)
• Matrix Solution (R2)
• Substrate Solution (R3)
• FeSO₄-7H₂O Standard
• Disposable 96-well microplate

The FRAP working solution is prepared by mixing the reagents in a 10:1:1 ratio prior to the assay.

Sample Preparation

The kit supports analysis of various sample types with minimal preparation:

• Serum / Plasma: centrifuge blood samples and collect supernatant
• Tissue Samples: homogenize tissue in PBS or saline and centrifuge
• Cells: disrupt cells via homogenization or sonication
• Saliva / Urine: direct analysis after centrifugation if necessary

These preparation methods ensure accurate release and measurement of antioxidant molecules in biological samples.

Detection Method

• Measurement wavelength: 593 nm
• Reaction time: 3–5 minutes at 37°C
• Suitable for microplate reader or spectrophotometer analysis.

Applications

The Total Antioxidant Capacity Assay Kit is widely used in research areas including:

• Oxidative stress and redox biology research
• Antioxidant activity analysis
• Metabolic and physiological studies
• Drug and natural compound antioxidant evaluation
• Biomedical and biochemical research

Storage Conditions

• Reagents should be stored at −20°C and protected from light.
• Prepared FRAP working solution should be used within 2 hours for optimal assay performance.

Important Notice

For Research Use Only (RUO).
This product is intended strictly for laboratory research and not for clinical diagnosis or therapeutic applications.