IRON/TIBC REAGENT SET
INTENDED USE
Iron/TIBC Reagent Set can be used for the quantitative determination of iron, total iron-binding capacity in human serum.
INTRODUCTION
The iron content of the human body may be divided into three classes: iron in storage, iron in use, and iron in transport. Iron in storage is reserved iron contained within the cells. Iron in use contained in hemoglobin, various enzymes, and several other types of proteins. Iron in transport is being moved to storage or is being removed from storage to be utilized in the formation of hemoglobin, etc. Iron in a free state is not only relatively insoluble, but it is toxic.
Therefore, nearly all iron in the body is attached to some type of protein. It is of fundamental importance to note that a specimen should be analyzed for both iron and iron binding capacity because of the need for both values in the differential diagnosis of various types of anemia and liver diseases. For this reason, the current procedure is designed for the simultaneous determination of iron and iron binding capacity.
Serum iron assays measure transport iron bound to the protein transferrin. Increase in serum iron levels may indicate increased erythrocyte destruction, decreased erythrocyte formation, increased absorption, or detects in storage capabilities. Decrease in serum iron levels may indicate iron deficiency or inability to retrieve storage iron. Iron binding capacity is usually increased in iron deficient anemia and decreased in hemochromatosis, malignancies, rheumatic fever, Hodgkin's diseases, and collagen vascular disease.
Most successful iron methodologies remove iron from transferrin, reduce it to the ferrous state, bind it to a chromophore, and quantitate it by measuring the amount of color developed. The determination of Iron binding capacity involves adding sufficient iron to saturate transferrin and then determining either the total amount of bound iron or the excess unbound iron. The later procedure is applied to determine Iron Binding Capacity.
PRINCIPLE
Iron/TIBC Reagent Set
The iron in serum is dissociated from its Fe (III) – transferring complex by the addition of an acidic buffer containing hydroxylamine. This addition reduces the Fe (III) to Fe (II). The chromogenic agent, Ferene, forms a highly colored Fe (II) – complex that is measured photometrically at 560 nm.
The unsaturated iron binding capacity (UIBC) is determined by adding Fe (II) iron to serum so that they bind to the unsaturated iron binding sites on transferrin. The excess Fe (II) ions are reacted with Ferrozine to form the color complex, which measured photometrically. The difference between the amount of Fe (II) added and the amount of Fe (II) measured represents the unsaturated iron binding. The total iron binding capacity (TIBC) is determined by adding the serum iron value to the UIBC value.
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