ELISA kit for detecting airborne allergen based on IgM antibody
An antibody detection and kit technology, which is applied in the field of allergen detection, can solve the problems of inability to timely and accurately determine allergic diseases and stay, and achieve the effect of improving sensitivity, various types and wide application range.
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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 a coating solution for the airborne allergen;
[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 test plate wells overnight in step 2), add blocking solution, and wash again after incubation to obtain a test plate coated with airborne allergens.
[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 carbo...
Embodiment 1
[0088] Coating: Dilute Humulus japonicus pollen and Artemisia grandis pollen allergens with carbonate buffer solution to airborne allergen coating solution with a mass concentration of 10 μg / ml; dilute human IgM protein with carbonate buffer solution to obtain standard products Coating solution (see Table 1 for protein concentration). Add 0.05ml of coating solution to each reaction well of the polystyrene plate, overnight at 4°C. On the next day, the solution in the wells was discarded, and 0.35ml of washing buffer was added to each well, washed 3 times, and incubated 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: sera from patients with suspected humulus pollen allergy, sera from patients with wormwood pollen allerg...
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