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Total Iron Colorimetric Assay Kit– MSE Supplies LLC

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Total Iron Colorimetric Assay Kit

SKU: E-BC-K772-M-96

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Total Iron Colorimetric Assay Kit

SKU # E-BC-K772-M
Detection Instrument Microplate reader (590-600 nm, optimum wavelength: 593 nm)
Detection method: Colorimetric method


Product Details

Properties

Sample type Serum, tissue
Sensitivity 0.4 μmol/L
Detection range 0.4-50 μmol/L
Detection method Colorimetric method
Assay type Quantitative
Assay time 80 min
Precision Average inter-assay CV: 1.500% | Average intra-assay CV: 1.300%
Storage 2-8℃
Valid period 12 months


Images

H Tian et al developed a targeted nanomodulator that can induced tumor cell ferroptosis. Iron (Fe) level in mouse tumor cells 4T1 were determined using total iron colorimetric assay kit (E-BC-K772-M).

The increased iron content in 4T1 cells showed significantly difference between different treatments.

 

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 (0.4-50 μmol/L).

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

Sample type Dilution factor
Human serum 1
Mouse serum 1-2
Rat serum 1
10% Mouse liver tissue homogenate 2-3
10% Rat lung tissue homogenate 1
10% Mouse heart tissue homogenate 1
10% Rat spleen tissue homogenate 2-3
10% Epipremnum aureum leaf tissue homogenate 1

 

Note: The diluent is reagent 1.

 

Detection Principle

Under the action of reductant, iron ions in samples can be reduced into ferrous ions (Fe2+). The latter then bind to probe and form complexes, which has a maximum absorption peak at 593 nm. The concentration of iron can be calculated by measuring the OD value at 593 nm indirectly.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Buffer Solution 50 mL × 1 vial 50 mL × 2 vials 2-8℃, 12 months,
shading light
Reagent 2 Chromogenic
Solution
10 mL × 1 vial 10 mL × 2 vials 2-8℃, 12 months,
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Reagent 3 10 mmol/L Iron Standard 1 mL × 1 vial 1 mL × 2 vials 2-8℃, 12 months,
shading light
Reagent 4 Extracting Solution 40 mL × 1 vial 40 mL × 2 vials 2-8℃, 12 months,
shading light
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 (μmol/L) 1.80 16.50 40.50
%CV 1.5 1.3 1.1


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 (μmol/L) 1.80 16.50 40.50
%CV 1.2 1.7 1.6


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 99%.


Standard 1 Standard 2 Standard 3
Expected Conc. (μmol/L) 7.5 18 36
Observed Conc. (μmol/L) 7.6 17.8 34.9
Recovery rate(%) 101 99 97

 

Sensitivity

The analytical sensitivity of the assay is 0.4 μmol/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 (μmol/L) 0 5 10 15 20 30 40 50
Average OD 0.048 0.136 0.232 0.326 0.417 0.611 0.795 0.978
Absoluted OD 0.000 0.088 0.184 0.278 0.369 0.563 0.747 0.930