Total IgE ELISA kit is an enzyme immunoassay (microtiter strips) for the quantitative determination of immunoglobulin E (IgE) in human serum and plasma. The lowest detectable level using this Total IgE ELISA kit that can be distinguished from the zero standard is 1.0 IU/ml (defined as mean of the ODs of the zero calibrator minus 2 SD) as read from the standard curve. The intra-assay precision (n=20) for this Total IgE ELISA kit is 4,8% to 6,5%, the inter-assay precision (n=23) was determined to be 5,6% to 8,7%. The recovery of IgE for this Total IgE ELISA kit is 90 - 110%. The Total IgE kit test was designed so, that there is no “high-dose hook effect” for the majority of elevated IgE concentrations. No “high-dose hook effect” has been observed up to 50,000 IU/ml. Samples with higher absorptions than the highest calibrator should however be diluted by the zero calibrator (e.g. 1:5) and re-assayed to obtain correct values.
The Total IgE ELISA kit provides materials for the quantitative measurement of immunoglobulin E (IgE) in human serum and plasma. This Total IgE ELISA kit assay is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. A monoclonal mouse-antihuman IgE antibody is bound on the surface of the microtiter strips. Patient samples or standards are pipetted into the wells of the microtiter plate together with ready to use anti-human IgE peroxidase conjugate. A sandwich-like binding between the antibodies of the serum, the immobilized antibody and the conjugate takes place. After a 30 minutes´ incubation at room temperature, the plate is rinsed with diluted wash solution, in order to remove unbound material. Then the substrate (TMB) solution is pipetted and incubated for 15 minutes, inducing the development of a blue dye in the wells. The colour development is terminated by the addition of a stop solution, which changes the colour from blue to yellow. The resulting dye is measured spectrophotometrically at the wavelength of 450 nm. The concentration of the antibodies is directly proportional to the intensity of the colour. The Total IgE ELISA kit is shipped at ambient temperature and should be stored at 2 – 8 °C. Once the foilbag of the coated microtiter strips has been broken, care should be taken to close it tightly again. The immunoreactivity of the coated microtiter strips of this Total IgE ELISA kit is stable up to 3 months in the broken, but tightly closed bag when stored at 2 – 8 °C.
The existence of IgE in man as a unique class of immunoglobulins which are important in the mediation of the allergic response has been known for over twenty years. The mechanism of action involves an initial antigenic stimulation of immunocompetent B lymphocytes by a specific antigen, a process which induces the lymphocyte to respond by producing specific antibody of several classes. One class, reaginic or IgE antibody, becomes partially bound via its Fc portion to receptors on the surface of mast cells and basophilic leukocytes. Upon further stimulation by specific allergens, these cellbound IgE molecules bind via their Fab portion to the allergen. This combination triggers the mast cells and basophilic leucocytes to release various vasoactive amines into the blood and the surrounding tissue. These substances cause smooth muscle constriction and lead ultimately to allergic conditions such as wheal and flare reactions, hives, dermatitis, rhinitis, hay fever, asthma and anaphylactic shock. IgE determinations are most valuable in the diagnostic assesment of patients with established or suspected allergic desease. In normal subjects, IgE values are related to age, with normal values peaking around 10 - 14 years. Infants and children with family history of atopic allergy are at increased risk of developing disease and constitute a prime population for screening. Studies have shown that conditions such as asthma, rhinitis, eczema, urticaria, dermatitis and some parasitic infections lead to increased IgE levels. Asthma, hay fever and atopic eczema patients may produce levels 3 - 10 times those of normal patients. Low IgE levels are found in hypogammaglobulinemia, and low or normal levels are associated with patients with autoimmune diseases, ulcerative colitis, infections, serum hepatitis, cancer and malaria.
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