IgE CLIA Kit

Catalog No. ABIN504803

Product Details IgE CLIA Kit

Target: IgE

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: Sandwich ELISA

Application: ELISA

Analytical Method: Quantitative

Characteristics: The Quantitative Determination of Immunoglobulin E (IgE) Concentration in Human Serum by a Microplate Chemiluminescence Immunoassay.

Application Details

Application Notes: All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.

Sample Volume: 25 μL

Plate: Pre-coated

Protocol:

Specimien Collection and Preparation:

The specimens shall be blood, serum in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot for samples. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2_x001E_8oC for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20oC for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.050ml of the specimen is re_x001F_quired.

Reagent Preparation:

1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store diluted buffer at room temperature 20-27(C. 2. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly.

Test Procedure:

Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27( C). 1. Format the microplates wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.025 ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of the IgE Biotin Reagent to each well. It is very important to dispense all reagents close to the bottom of the coated well. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 30 minutes at room temperature. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 7. Add 350l of wash buffer (see Reagent Preparation Section) decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. 8. Add 0.100 ml (100l) of the IgE Tracer Reagent to each well. DO NOT SHAKE THE PLATE AFTER TRACER ADDITION. 9. Incubate 30 minutes at room temperature. 10. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 11. Add 350l of wash buffer (see Reagent Preparation Section) decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. 12. Add 0.100 ml (100l) of working signal reagent to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time. 13. Incubate at room temperature for five (5) minutes in the dark. 14. Read the Relative Light Units (RLU) in each well for 0.2 1.0 seconds. The results should be read within thirty (30) minutes of adding the stop solution.

Restrictions: For Research Use only

Handling

Target Details

Target: IgE

Abstract: IgE Products

Synonyms: IGH2 CLIA Kit, IgE CLIA Kit, Gm900 CLIA Kit, immunoglobulin heavy chain (epsilon polypeptide) CLIA Kit, immunoglobulin heavy constant epsilon CLIA Kit, Immunoglobulin heavy constant epsilon CLIA Kit, Ighe CLIA Kit, IGHE CLIA Kit

Background: Allergic reactions, which are becoming more widespread, are usually diagnosed on the basis of medical history and clinical symptoms. In vitro and in vivo testing, however, play a key role in confirming clinical suspicions and tailoring treatment. The measurement of immunoglobulin E (IgE) in serum is widely used in the diagnosis of allergic reactions and parasitic infections. Many allergies are caused by the immunoglobulins of subclass IgE acting as point of contact between the allergen and specialized cells. The IgE molecules (MW 200,000) bind to the surface of the mast cells and basophillic granulocytes. Subsequently the binding of allergen to cell-bound IgE causes these cells to release histamines and other vasoactive substances. The release of histamines in the body results initiates what is commonly known as an allergic reaction. Before making any therapeutic determination it is important, however, to know whether the allergic reaction is IgE mediated or non-IgE mediated. Measurement of total IgE in serum sample, along with other supporting diagnostic information, can help to make that determination. Measurement of total circulating IgE may also be of value in the early detection of allergy in infants and as a means of predicting future atopic manifestations. Before deciding on any therapy it is important to take into consideration all the relevant clinical information as well as information supplied by specific allergy testing. IgE levels show a slow increase during childhood, reaching adult levels in the second decade of life. In general, the total IgE levels increase with the allergies a person has and the number of times of exposure to the relevant allergens. Significant elevations may be seen in the sensitized individuals, but also in cases of myeloma, pulmonay aspergillosis, and during the active stages of parasitic infections. In this method, IgE calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for IgE) is added and the reactants mixed. Reaction between the IgE antibodies and native IgE forms complex that binds with the streptavidin coated to the well. The excess serum proteins are washed away via a wash step. Another enzyme labeled monoclonal antibody specific to IgE is added to the wells. The enzyme labeled antibody binds to the IgE already immobilized on the well through its binding with the biotinylated monoclonal antibody. Excess enzyme is washed off via a wash step. A light signal is generated by the addition of a substrate. The intensity of the light generation is directly proportional to the concentration of the IgE in the sample. _x000E_