Do you know the experimental principle of the ELISA kit? -Huaqiang Electronic Network

When friends are just getting into the ELISA experiment, they must first understand what the principle is, so today Shanghai Huding talks about the experimental principle of the elisa kit. The kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards of known concentration and samples of unknown concentration are added to the microplates for detection. The biotinylated antibody is first incubated simultaneously. After washing, avidin-labeled HRP was added. After incubation and washing, the unbound enzyme conjugate is removed, then the substrates A, B, and the enzyme conjugate are added simultaneously. Produce color. The depth of the color is proportional to the concentration in the sample.
Self-contained materials
1. Distilled water.
2. Sampler: 5ul, 10ul, 50ul, 100ul, 200ul, 500ul, 1000ul.
3. Oscillators and magnetic stirrers, etc.
safety
1. Avoid direct contact with the stop solution and substrates A, B. Once exposed to these liquids, rinse with water as soon as possible.
2. Do not eat, drink, smoke or use cosmetics during the experiment.
3. Do not use your mouth to take any ingredients from the kit.
4. Reagents should be stored according to the label instructions and returned to room temperature before use. Standards after dilution should be discarded and cannot be stored.
5. The slats not used in the experiment should be immediately put back into the bag and sealed to prevent deterioration.
6. Other reagents not used should be packaged or covered. Do not mix reagents of different batches. Use before warranty.
7. Use a disposable tip to avoid cross-contamination. Avoid pipettes with metal parts when drawing stop solution and substrate A and B.
8. Use a clean plastic container to configure the wash solution. Mix all the ingredients and samples in the kit thoroughly before use.
9. Wash the enzyme plate should be fully patted dry, do not put the absorbent paper directly into the enzyme standard reaction well to absorb water.
10. Substrate A should be volatilized to avoid opening the lid for extended periods of time. Substrate B is sensitive to light and avoids prolonged exposure to light. Avoid contact with hands and be toxic. The OD value should be read immediately after the experiment is completed.
11. The order of addition of reagents should be consistent to ensure that all wells are incubated for the same time.
12. The incubation was carried out according to the time indicated in the instructions, the amount of addition and the order.
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