Weiwei Bio for you to explain indirect method to test antibodies - Database & Sql Blog Articles

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 it is called an indirect method. The steps are as follows:

one,

The specific antigen is linked to a solid phase carrier to form a solid phase antigen. Washing removes unbound antigen and impurities.

two,

Add the diluted test serum and incubate the reaction. The specific antibody in serum binds to the solid phase antigen to form a solid phase antigen antibody complex

Compound. After washing, only specific antibodies are left on the solid support, and other components in the serum are washed away during the washing process.

three,

Add enzyme-labeled anti-antibody. Anti-human Ig can be used to detect total antibodies, but IgG anti-antigen is generally detected by enzyme-labeled anti-human IgG.

body. The antibody in the solid phase immune complex binds to the enzyme-labeled antibody, thereby indirectly labeling the enzyme. After washing, solid phase loading

The amount of enzyme on the body is positively correlated with the amount of antibody tested in the specimen.

four,

Add substrate color

This method is mainly used for the diagnosis of infectious diseases for the detection of pathogen antibodies. The advantage of the indirect method is that a method for detecting the corresponding antibody can be established by using the same enzyme-labeled anti-antibody as long as the transformation is coated with the antigen.

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