FRAP Antioxidant Capacity Assay Kit

Catalog No. ABIN7384576

Product Details

Detection Method: Colorimetric

Detection Range: 7.5-250 μg/mL

Minimum Detection Limit: 7.5 μg/mL

Application: Biochemical Assay (BCA)

Purpose: The kit is designed for the antioxidant capacity measurement in biological samples.

Specificity: Specific for antioxidant activity determination.

Characteristics: FRAP assay kit is recommended for total antioxidant activity of single antioxidants in aqueous solution and added to plasma. The assay described here measures the ferric reducing ability of plasma (FRAP). At low pH , when a ferric complex is red

Components: Plate and the reagents neccesary to perform the assay.

Material not included: Pipettes, reaction tubes, plate reader.

Application Details

Sample Volume: 10 μL

Assay Time: 15 min

Plate: Uncoated

Reagent Preparation:

Reagents Preparation: 1. Add exactly 3.5 mL of ultrapure water in each vial of Reagent B and mix thoroughly. Once dissolved, keep refrigerated at -20 °C. (solution B). 2. Prepare FRAP working solution just before use by mixing Reagents A, B and C (10:1:1). 3. FRAP standard: Add exactly 1 mL of ultrapure water in each Standard vial and mix thoroughly. Prepare standards immediately prior to the assay performed. Do not store the standard preparations.

Sample Preparation:

Sample Preparation: Dilute your sample to an absorbance value corresponding to 500-600 μM of standard approximately. Plasma samples do not usually need to be diluted.

Assay Procedure:

Standard Preparation Antioxidant activity is expressed as FRAP values (Ferric Reducing Ability of Plasma). These values are related to Fe2+ concentration. Prepare calibration curve in 1 mL tubes as shown below in Table 1 (see kit booklet).Performing the assay: 1. Add 10 μL of the sample or standard in each well. 2.Add 220 μL of FRAP working solution previously prepared (see Reagents Preparation) in each well. Final dilution of the sample in the reaction mixture will be therefore, 1/22. 3.Mix the mixture for 4 minutes under continuous stirring. 4.Read the absorbance at 593 nm.

Calculation of Results:

1.Zeroed the absorbance values: ?A593nm = ?A593nm sample/standard - A593nm blank. Where A593nm sample/standard is the absorbance measured 4 minutes after the addition of antioxidants from samples or standards. 2.Plot the zeroed absorbance (?A593nm) of standards as a function of their final concentrations (Table 1). See Figure 2 for a typical standard curve (see images). 3.Calculate the FRAP value of the samples using the equation obtained from the linear regression of the standard curve substituted ?A593nmvalues for each sample. FRAP (μM) = (?A593nm- intercept)/ slope

Restrictions: For Research Use only

Handling

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C