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Total Antioxidant Capacity (T-AOC) Colorimetric Assay Kit(FRAP Method)– MSE Supplies LLC

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Total Antioxidant Capacity (T-AOC) Colorimetric Assay Kit (FRAP Method)

SKU: E-BC-K225-M-500

  • $ 40895
  • Save $ 7200



Total Antioxidant Capacity (T-AOC) Colorimetric Assay Kit (FRAP Method)

SKU # E-BC-K225-M
Detection Instrument Microplate reader (590-600 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms T-AOC
Sample Type Serum, plasma, saliva, urine, tissue, cells, cell culture supernatant
Sensitivity 0.049 mmol/L
Detection Range 0.049-2.5 mmol/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 20 min
Precision Average inter-assay CV: 8.100% | Average intra-assay CV: 3.900%
Other instruments required Balance, Incubator, Micropipettor
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
Storage 2-8℃
Valid period 12 months


Images

D Shanmugarajan et al investigate the antihypertensive and pleiotropic effect of Phyllanthus emblica extract. Total antioxidant capacity (T-AOC) in human serum was determined using T-AOC colorimetric assay kit (E-BC-K225-M).

No significant difference in total oxidant status between Phyllanthus emblica group and placebo group can be seen.

 

Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.049-2.5 mmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Human serum 1
Human saliva 1
Human urine 1
Cellular supernatant 1
HepG2 cells homogenization 1
5% Mouse liver tissue homogenization 1
10% Epipremnum aureum tissue homogenization 1

 

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

 

Detection Principle

Fe3+-TPTZ can be reduced by antioxidants and produce blue Fe2+-TPTZ under acid condition. The antioxidant capacity of sample can be calculated by detection the absorbance value at 593 nm.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Buffer Solution 10 mL × 1 vial 20 mL × 1 vial 2-8℃, 12 months
Reagent 2 TPTZ Solution 1 mL × 1 vial 2 mL × 1 vial 2-8°C, 12 months, shading light
Reagent 3 Substrate Solution 1 mL × 1 vial 2 mL × 1 vial 2-8℃, 12 months
shading light
Reagent 4 FeSO4•7H2O
Standard
100 mg × 1 vial 200 mg × 1 vial 2-8℃, 12 months
Microplate 96 wells No requirement
Plate Sealer 2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters Sample 1 Sample 2 Sample 3
Mean (mmol/L) 0.58 1.02 1.95
%CV 4.3 4.0 3.4


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (mmol/L) 0.58 1.02 1.95
%CV 7.6 8.4 8.3

 

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 103%.

Standard 1 Standard 2 Standard 3
Expected Conc. (mmol/L) 0.5 1.1 2
Observed Conc. (mmol/L) 0.5 1.2 2.1
Recovery rate (%) 101 105 103

 

Sensitivity

The analytical sensitivity of the assay is 0.049 mmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

 

Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (mmol/L) 0 0.3 0.6 0.9 1.2 1.8 2.1
Average OD 0.070 0.160 0.253 0.356 0.423 0.618 0.708
Absoluted OD 0 0.090 0.184 0.287 0.354 0.548 0.639