antibody  (IgE) ELISA Test Kit

INTENDED USE

  This kit is a quantitative detection of total IgE antibody (IgE) in human serum/plasma in vitro. This result can assist type Ⅰ allergy diagnosis. Moreover, total IgE testing may also be recommended for patients with suspected parasitic diseases.

PRINCIPLE OF THE TEST

  Immediate hypersensitivity (typeⅠ allergies) is mediated by specific IgE. IgE concentration in normal serum peaks between 6 and 15 years of age. In most cases, the increase of specific IgE in patients is accompanied by the increase of total IgE titer. In this case, the titer can go up to 1,000 times. In general, international units per milliliter (IU/mL) are defined as: 1 IU/mL equals 2.4ng IgE. In patients with hereditary allergic dermatitis, IgE levels can be as high as 50,000 IU/mL. In addition, IgE titers are elevated in patients with parasitic diseases. Higher-than-normal test results should also take into account the impact of confirmed autoimmune diseases.

The reagent is not recommended for healthy people physical examination, testing results of positive or negative only on behalf of the corresponding IgE antibody test results positive or negative, uncertainty correlation with patient is sick and may not be as the only index of evaluation of patients, must be combined with patient clinical presentation and other laboratory tests for integrated analysis of the illness.

The detection method of this kit is enzyme-linked immunoassay. In this reagent, the principle of double antibody sandwich immunoassay was adopted. Monoclonal antibodies that recognize anti-IgE were pre-coated in the wells of the microplate, and human serum/plasma samples and enzyme-labeled another monoclonal antibody were added into the pores of the plate. After incubation and washing, substrate solution was added. OD value was read and its content was calculated by a microplate reader.

TEST PROCEDUR

1.All reagents should be allowed to reach room temperature for 15 minutes before use.

2. Dilute the wash buffer at the rate of 1:40 dilution with distilled water before use.

3. The sample should be corresponding to the number of micro plate, each plate should be provided with reference wells (The number of well is determined by reference and specimen to be tested).

Note: Use a separate disposal pipette tip for each specimen, each reference as to avoid cross contamination.

4.Add 100μL Sample diluent in the corresponding well, add 20μL each reference S0, S1, S2, S3, S4, S5 and sample respectively to corresponding well.

5. Shake gently to mix. Incubate at 37°C for 30 minutes with the sealing plate membrane sealing the plate.

6. At the end of the incubation, remove and discard the plate cover. Take out, add wash buffer to each well for 10 seconds. Repeat 5 times. After the final washing cycle, turn the plate over onto blotting paper or clean towel, and tap it to remove any remainders.

7. Respectively adding Conjugate 100µL. Incubate at 37°C for 30 minutes with the sealing plate membrane sealing the plate.

8. Wash the plate as the step 6.

9. Add Substrate A (50µL) and Substrate B (50µL). Shake gently to mix. Incubate at 37°C for 10 minutes with the sealing plate membrane sealing the plate.

10. Add 50μL Stop Solution to each well. Mix gently by shaking, read the absorbance within 10 minutes after stopping the reaction. Then read the absorbance value at 450nm/ 630nm with the microplate reader. Calculate the concentration and evaluate the results.