ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting airborne allergen based on IgG2 antigen
A technology of antibody detection and kit, which is applied in the field of airborne allergen-specific IgG2 detection kit, which can solve the problems of inability to timely and accurately determine allergic diseases and retention, and achieve high accuracy, variety, and reproducibility Good results
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[0048] In the present invention, the preparation method of the detection plate coated with airborne allergens preferably includes the following steps:
[0049] 1) diluting the airborne allergen with a coating buffer to obtain the airborne allergen coating solution;
[0050] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;
[0051] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.
[0052] In the invention, the coating buffer is used to dilute the airborne allergen to obtain the coating solution. In the present invention, the dilution method is not particularly limited, and a dilution technique known to those skilled in the art can be used. In the present invention, the coating buffer is preferably carbonate buffer. In the present invention, the concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L, more pr...
Embodiment 1
[0088]Coating: Dilute dust mite, Artemisia grandis pollen and Humulus pollen allergens with carbonate buffer solution to the airborne allergen coating solution with a mass concentration of 10 μg / ml; dilute human IgG2 protein with carbonate buffer solution Obtain standard coating solution (protein concentration is shown in Table 1). Add 0.05ml of coating solution to each reaction well of the polystyrene plate, overnight at 4°C. On the next day, discard the solution in the well, add 0.35ml washing buffer to each well, wash 3 times, and incubate for 0.5min each time (referred to as washing, the same below);
[0089] Blocking: Add 0.05ml of PBS buffer solution containing 2% bovine serum albumin to each reaction well, pH: 7.2 (blocking solution), incubate at 15-35°C for 120min, discard the solution in the well, and wash as above;
[0090] Add samples: serum from patients with suspected dust mite allergy, serum from patients with Artemisia grandis pollen allergy, serum from patient...
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