Microporous plate and kit for food allergen IgG antibody detection, preparation methods thereof, and detection method
A technology for food allergy and antibody detection, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of time-consuming, labor-intensive, expensive, and the cause of disease cannot be found, and achieve considerable economic benefits, high costs, and good social benefits.
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Embodiment 1
[0038] Example 1: Preparation of 14 kinds of food allergen IgG antibody detection microwell plates and kits
[0039] 1. Dissolve 14 food allergen proteins (beef, chicken, cod, corn, crab, egg white / yolk, mushroom, milk, pork, rice, shrimp, soybean, tomato, wheat) in 50mM carbonic acid buffer at 10μg / ml solution, mix well.
[0040] 2. Put them into the microwell plate in a certain order (100 μl per well), 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), 200 μl per well, and incubated overnight at 4°C.
[0041] 3. Discard the liquid in the well, dry the microporous plate overnight at room temperature, and seal the microporous plate in an aluminum foil bag after drying.
[0042] 4. Weigh 10g of BSA and dissolve it in 1 liter of 50mM phosphate buffer solution to prepare sample diluent, and pack in 50ml bottles.
[0043] 5. Horseradish peroxidase and anti-human Ig...
Embodiment 2
[0052] Embodiment 2: the detection method of food allergen IgG antibody
[0053] Equilibrate all reagents to room temperature before use.
[0054] 1. Prepare to draw a standard curve:
[0055] Take four 12x75mm glass tubes and mark 50, 100, 200 and 400U / ml respectively. Add 150 μl of serum dilution to these 4 tubes sequentially. Add 150 μl of 14 kinds of food allergen IgG antibody standard serum to the tube marked 400U / ml. After mixing, take 150μl and add it to the tube marked 200U / ml; after mixing again, take 150μl and add it to the tube marked 100U / ml; after mixing the same, take 150μl and add it to the tube marked 50U / ml. In this way, 150 μl of IgG antibody standard serum with a concentration of 100, 200 and 400 U / ml, respectively, and 300 μl of IgG antibody standard serum with a concentration of 50 U / ml were obtained. Take 100 μl from each tube and add it into the microwell plate: draw the standard curve with the concentration as the abscissa and the absorbance as the ...
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