The indirect method is a commonly used method for detecting antibodies. The principle is to use an enzyme-labeled anti-antibody (anti-human immunoglobulin antibody) to detect a test antibody that binds to a solid phase antigen, so the indirect method is as follows:
1) A specific antigen is linked to a solid phase carrier to form a solid phase antigen. Washing removes unbound antigen and impurities.
2) Add diluted test serum and incubate the reaction. The specific antibody in the serum binds to the solid phase antigen to form a solid phase antigen-antibody complex. After washing, only specific antibodies are left on the solid support, and other components in the serum are washed away during the washing process.
3) Add enzyme-labeled anti-antibody. The antibody can be labeled with an anti-human Ig to detect total antibodies, but IgG antibodies are generally detected by using an enzyme-labeled anti-human IgG. The antibody in the solid phase immune complex binds to the enzyme-labeled antibody, thereby indirectly labeling the enzyme. After washing, the amount of enzyme on the solid support is positively correlated with the amount of antibody tested in the specimen.
4) Adding substrate color
Human ciliary neurotrophic factor (CNTF) ELISA kit specimen collection notes: 1) There must be a complete plan before collecting the specimen. It must be clear whether the ingredients to be tested are stable enough.
2) We advocate the early detection of fresh specimens and timely detection after collection.
1. Detection principle: double antibody sandwich ABC-ELISA
2. The kit is stored at 2-8 ° C (when used frequently); -20 ° C (when not used for a long time).
3. The newly opened enzyme-linked plate well may contain a little water-like substance, which is normal and will not have any effect on the experimental results.
4. Chinese and English manuals may be inconsistent. Please refer to the English manual.
5. When the concentrated washing liquid is stored at a low temperature, salt will be precipitated, and when diluted, it can be heated and dissolved in a water bath.
6. Do not replace the reagents in the kit with reagents from other manufacturers.
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