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|Method & Line||Sample & Target||Product||Package Info||Product||Package Info|
|MicroELISA||Serum||EIAgen Ferritin Kit||Tests per Package: 96|
|EIAgen||The EIAgen Ferritin Kit has been designed for the quantitative determination of Circulating Ferritin Concentrations in Human Serum.||Code: LI4023||Package: 1 Microplate|
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The EIAgen Ferritin Kit has been designed for the quantitative determination of Circulating Ferritin Concentrations in Human Serum.
Ferritin, in circulation, as measured in serum levels is a satisfactory index of body’s iron storage. The iron storage is directly measured by quantitative phlebotomy, iron absorption studies, liver biopsies and microscopic examinations of bone marrow aspirates. Iron deficiency (Anemia) and iron overload (Hemochromatosis) are conditions associated with body’s iron storage or lack thereof. Measurements of total iron binding capacity (TIBC) have widely been used as aids in the determination of these conditions. However, an assay of serum Ferritin is simply more sensitive and reliable means of demonstration these disorders. Ferritin is present in blood in very low concentrations. Normally, approximately 1% of plasma iron is contained in Ferritin. The plasma ferritin, is in equilibrium with body stores, and variations of iron storage. The plasma concentrations of ferritin decline very early in anemic conditions like development of iron deficiency, long before the changes are observed in the blood hemoglobin concentration, size of the erythrocytes and TIBC. Thus measurements of serum ferritin can serve as an early indicator of iron deficiency that is uncomplicated by other concurrent conditions. At the same time a large number of chronic conditions can result in elevated levels of serum ferritin. These include chronic infections, chronic inflammatory diseases such as rheumatoid arthritis, heart disease and some other malignancies, especially lymphomas, leukemia, breast cancer and neuroblastoma. In patients who have these chronic disorders together with iron deficiency, serum ferritin levels are often normal. An increase in circulating ferritin is observed in patients with viral hepatitis or after a toxic liver injury as a release of ferritin from the injured liver cells. Elevated serum ferritin levels are found in patients with hemochromatosis and hemosiderosis. Circulating ferritin levels have been used by clinicians, as an aid, in the diagnosis of several other disorders. It has proved as a valuable tool in differential diagnosis of anemia due to iron deficiency and anemias due to other disorders and, in exposing the depletion of iron reserves long before the onset of anemia. Serial determinations have been used to monitor, non-invasively, the erosion of iron storage during pregnancy and in patients undergoing dialysis. Serum ferritin is routinely used as a screen for iron deficiency for a variety of populations like blood donors and people who are receiving regular blood transfusions or iron replacement therapy. In this method, ferritin calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for ferritin) is added and the reactants mixed. Reaction results between the biotinylated ferritin antibody and native ferritin to form an immune complex that is deposited on the streptavidin coated well. The excess serum proteins are washed away via a wash step. Another ferritin specific antibody, labeled with an enzyme, is added to the wells. The enzyme labeled antibody binds to the ferritin already immobilized on the well. Excess enzyme is washed off via a wash step. A color is generated by the addition of a substrate. The intensity of the color generation is directly proportional to the concentration of the ferritin in the sample. The employment of several serum references of known ferritin levels permits the construction of a dose response curve of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with ferritin concentration.
Immunoenzymometric sequential assay (TYPE 4): The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme and immobilized), with different and distinct epitope recognition, in excess, and native antigen. In this procedure, the immobilization takes place during the assay at the surface of a microplate well through the interaction of streptavidin coated on the well and exogenously added biotinylated monoclonal anti- ferritin antibody.
Upon mixing monoclonal biotinylated antibody, and a serum containing the native antigen, reaction results between the native antigen and the antibody, forming an antibody-antigen complex. Simultaneously the biotin attached to the antibody binds to the streptavidin coated on the microwells resulting in immobilization of the complex.
After a suitable incubation period, the antibody-antigen bound fraction is separated from unbound antigen by decantation or aspiration. Another antibody (directed at a different epitope) labeled with an enzyme is added. Another interaction occurs to form an enzyme labeled antibody-antigen-biotinylated-antibody complex on the surface of the wells. Excess enzyme is washed off via a wash step. A suitable substrate is added to produce color measurable with the use of a microplate spectrophotometer. The enzyme activity on the well is directly proportional to the native antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
The kit contains reagents for 96 tests (code LI4023).
Bring reagents to room temperature before use.
Substrate A (TMB)
Substrate B (H2O2)
Stop Solution (HCl - 1N)
Wash Buffer Conc. 50X
Number of tests