Microporous plate and kit for food allergen IgG antibody detection, preparation methods thereof, and detection method

[0038] Example 1: Preparation of 14 kinds of food allergen I gG antibody detection microplate and kit

[0039] 1. Dissolve 14 food allergen proteins (beef, chicken, cod, corn, crab, egg white / yolk, mushrooms, milk, pork, rice, shrimp, soybeans, tomatoes, wheat) in 50 mM carbonate buffer at 10 μg / ml In the liquid, mix well.

[0040] 2. Add to the microtiter plate (100μl per well) in a certain sequence, incubate overnight at 4°C, wash the plate with Tween-containing phosphate buffer for 3 times, and then add blocking solution (containing 2 %BSA in phosphate buffer), 200μl per well, incubated overnight at 4°C.

[0041] 3. Discard the liquid in the well, dry the microplate overnight at room temperature, and seal the microplate in an aluminum foil bag after drying.

[0042] 4. Weigh 10g of BSA and dissolve it in 1 liter of 50mM phosphate buffer to prepare a sample diluent, and divide it into 50ml bottles.

[0043] 5. The horseradish peroxidase and anti-human IgG antibody (3:1) were labele

[0052] Example 2: Detection method of food allergen IgG antibody

[0053] Please equilibrate all reagents to room temperature before use.

[0054] 1. Prepare to draw a standard curve:

[0055] Take 4 12x75mm glass tubes and mark them as 50, 100, 200 and 400 U / ml. Add 150μl of serum diluent to these 4 tubes in sequence. Add 150μl of 14 kinds of food allergen IgG antibody standard serum to the 400U / ml tube. After mixing, take 150μl and add it to the tube labeled 200U / ml; after mixing again, take 150μl and add it to the tube labeled 100U / ml; after the same mixing, take 150μl and add it to the tube labeled 50U / ml. In this way, 150μl of IgG antibody standard serum with a concentration of 100, 200 and 400U / ml respectively, and 300μl of IgG antibody standard serum with a concentration of 50U / ml were obtained. Take 100μl from each tube and add it to the microplate: draw a standard curve with the concentration as the abscissa and the absorbance as the ordinate, and calculate th