Enzyme Immunoassay for the Quantitative Determination of Immunoglobulin E (IgE) Concentration in Human Serum

Product Description
Patients with atopic allergic diseases such as atopic asthma, atopic dermatitis, and hay fever have been shown to exhibit increased total immunoglobulin E (IgE) levels in blood. IgE is also known as the reagenic antibody.
In general, elevated levels of IgE indicate an increased probability of an IgE-mediated hypersensitivity, responsible for allergic reactions. Parasitic infestations such as hookworm, and certain clinical disorders including aspergillosis, have also been demonstrated to cause high levels of IgE.
Decreased levels of IgE are found in cases of hypogammaglobulinemia, autoimmune diseases, ulcerative colitis, hepatitis,cancer, and malaria. Cord blood or serum IgE levels may have prognostic value in assessing the risk of future allergic conditions in children.
The IgE serum concentration in a patient is dependent on both the extent of the allergic reaction and the number of different allergens to which he is sensitized.
Nonallergic normal individuals have IgE concentrations that vary widely and increase steadily during childhood, reaching their highest levels at age 15 to 20, and thereafter remaining constant until about age 60 when they slowly decline.

Principle
The IgE Quantitative Test is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay system utilizes one monoclonal anti-IgE antibody for solid phase (microtiter wells) immobilization and goat anti-IgE antibody in the antibody-enzyme (horseradish peroxidase) conjugate solution.
The test specimen (serum) is added to the IgE antibody coated microtiter wells and incubated with the Zero Buffer at room temperature for 30 minutes.
If human IgE is present in the specimen, it will combine with the antibody on the well. The well is then washed to remove any residual test specimen, then washed to remove any residual test specimen, and IgE antibody labeled with horseradish peroxidase (conjugate) are added.
The conjugate will bind immunologically to the IgE on the well, resulting in the IgE molecules being sandwiched between the solid phase and enzyme-linked antibodies. After incubation at room temperature for 30 minutes, the wells are washed with water to remove unboundlabeled antibodies.
A solution of TMB Reagent is added and incubated at room temperature for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution, and the color is changed to yellow and measured spectrophotometrically at 450nm.
The concentration of IgE is directly proportional to the color intensity of the test sample.

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