Manager talks online about elisa kits - Database & Sql Blog Articles

The day before, Mr. Xia of Southwest University purchased the elisa kit in our company after telephone consultation. On the phone, Mr. Xia and her manager, Mr. He, conducted an online Q&A:
Teacher Xia: How to deal with serum samples? Manager: The whole blood sample should be placed at room temperature for 2 hours or after 4 °Cguo night and centrifuged at 1000 × g for 20 minutes. Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
Teacher Xia: How to deal with plasma samples? Manager: EDTA, sodium citrate or heparin should be selected as anticoagulant according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge at 1000×g for 15 minutes, and collect the supernatant. If a precipitate forms during storage, it should be centrifuged again. Teacher Xia: How to treat cell supernatant samples? Manager: When detecting secreted ingredients, collect them with sterile tubes. Centrifuge for about 20 minutes (1000 x g). Collect the supernatant carefully. When the components in the cells were detected, the cell suspension was diluted with PBS (pH 7.2-7.4), and the cell concentration reached about 1 million/ml. By repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for about 10 minutes (5000 x g). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again. Teacher Xia: Why are some kits using competitive ELISA? Manager: Small molecule antigens or haptens cannot be measured by the double antibody sandwich method because they lack two or more sites that can be used as sandwich methods. Competition methods can be used. The principle is that the antigen in the specimen and the antigen on the solid phase carrier compete for the binding site on the biotinylated antibody, and the more the antigen content in the sample, the less biotinylated antibody bound to the solid phase, and the final color development. The shallower, that is, the color depth is inversely proportional to the antigen concentration in the sample. This method is often used for ELISA assays such as small molecule hormones and drugs.
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