Calbiotech Ferritin ELISA

SUMMARY AND EXPLANATION

Human Ferritin is a large molecule with a molecular weight of approximately 450,000 Daltons, and consists of a protein shell around an iron core; each molecule of Ferritin may contain as many as 4,000 iron atoms. Under normal conditions, this may represent 25% of the total iron found in the body. In addition, Ferrritin can be found in several isomers. High concentrations of Ferritin are found in the cytoplasm of the reticuloendothelial system, the liver, spleen and bone marrow. Methods previously used to measure iron in such tissues are invasive, cause patient trauma and lack adequate sensitivity. The measurement of Ferritin in serum is useful in determining changes in body iron storage, and is non-invasive with relatively little patient discomfort. Serum Ferritin levels can be measured routinely and are particularly useful in the early detection of iron-deficiency anemia in apparently healthy people. Serum Ferritin measurements are also clinically significant in the monitoring of the iron status of pregnant women, blood donors, and renal dialysis patients. High Ferritin levels may indicate iron overload without apparent liver damage, as may be noted in the early stages of idiopathic hemochromatosis. Ferritin levels in serum have also been used to evaluate clinical conditions not related to iron storage, including inflammation, chronic liver disease, and malignancy.

PRINCIPLE OF THE TEST 

This Ferritin ELISA kit is a solid phase sandwich assay method, based on a streptavidin-biotin principle. The standards, samples and the biotinylated Anti-Ferritin antibody reagent are added into designated wells, coated with Streptavidin. Endogenous Ferritin in the patient’s serum binds to the antigenic site of the biotinylated Anti-Ferritin antibody. Simultaneously, the biotinylated antibody is immobilized onto the wells through the high affinity Streptavidin-Biotin interaction. Unbound protein and excess biotin conjugated antibody are washed off by wash buffer. Upon the addition of the Peroxidase (HRP) conjugated AntiFerritin antibody reagent, a sandwich complex is formed, the analyte of interest being in between the two highly specific antibodies, labeled with Biotin and HRP. Unbound protein excess enzyme conjugated antibody reagent is washed off by wash buffer. Upon the addition of the substrate, the intensity of color developed is directly proportional to the concentration of Ferritin in the samples. A standard curve is prepared relating color intensity to the concentration of the Ferritin.