Iron Colorimetric Assay Kit

Iron Colorimetric Assay Kit

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C Pan et al investigate risk of microcytic anemia through chronic exposure to MC-LR. Iron (Fe) level in mouse serum and tissue were determined using iron colorimetric assay kit (E-BC-K139-S).

The serum iron levels were decreased in all MC-LR treatment groups (A), mice with MC-LR exposure developed iron overload in different tissue (B-F).

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 (0.072-60 mg/L).

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

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

Detection Principle

Under the action of acidic solution and reductant, ferric ions can be separated from transferrin in serum, and reduced into ferrous ions (Fe2+). The latter then bind to bipyridine and form pink complexes. The concentration of iron can be calculated by measuring the OD value at 520 nm indirectly.

Kit Components & Storage

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)

Inter-assay Precision

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

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%.

Sensitivity

The analytical sensitivity of the assay is 0.072 mg/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.