Immunoglobulin A (IgA) ELISA Kit
Basic Information
The content in normal human serum is second only to IgG, accounting for 10-20% of serum immunoglobulin content. From the structural point of view, IgA has a monomer, a dimer, a tris, and a multimer. According to its immune function, it is divided into two types: serotype and secretory. Serotype IgA is present in serum and accounts for about 85% of total IgA. Although serotype IgA has certain functions of IgG and IgM, it does not show important immune function in serum. Secretory IgA is present in secretions such as saliva, tears, colostrum, nasal and bronchial secretions, gastrointestinal fluids, urine, sweat, and the like. Secretory IgA is the main anti-infective immune antibody in the mucosa of the body. Therefore, it is also called mucosal local antibody. IgA cannot pass through the placenta. There is no IgA antibody in the serum of newborns, but secretory IgA can be obtained from breast milk. After 4 to 6 months of neonatal birth, IgA can appear in the blood, and then gradually increase, reaching a peak in adolescence.
Immunoglobulin A (IgA) ELISA Kit
Introduction to inspection technology
Double antibody sandwich method
1. Coat: The antibody was diluted to a protein content of 1 to 10 μg/ml with 0.05 MPH 9. 牰 carbonate coating buffer. 0.1 ml was added to the reaction well of each polystyrene plate at 4 ° C overnight. The next day, the solution in the well was discarded and washed 3 times with washing buffer for 3 minutes each time. (referred to as washing, the same below).
2. Loading: Add 0.1 ml of the sample to be tested to the above-mentioned coated reaction well, and incubate at 37 ° C for 1 hour. Then wash. (Do blank holes, negative control wells and positive control wells at the same time).
3. Add enzyme-labeled antibody: Add 0.1 ml of freshly diluted enzyme-labeled antibody (diluted titration) to each well. Incubate at 37 ° C for 0.5 to 1 hour and wash.
4. Add substrate solution to develop color: Add 0.1 ml of temporarily prepared TMB substrate solution to each reaction well at 37 ° C for 10 to 30 minutes.
5. Stop the reaction: Add 0.05 ml of 2M sulfuric acid to each reaction well.
6. Judgment of results: The results can be observed directly on the white background with the naked eye: the darker the color in the reaction well, the stronger the positive degree, the negative reaction is colorless or very light, according to the depth of the color, with "+", The "-" sign indicates. The OD value can also be measured: on the ELISA detector, at 450 nm (if the color is developed by ABTS, 410 nm), the OD value of each well is measured after zero adjustment of the blank control well, if it is greater than 2.1 times the OD value of the specified negative control. Is positive.
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