Hengyuan popular competition inhibition ELISA - Database & Sql Blog Articles

The competitive inhibition ELISA is also called blocking ELISA. The main principle is to use the antigen or antibody to be tested to interfere with the pre-designed system, and the final coloration result is to be tested. The degree of interference with antigen or antibody is inversely related. Competitive inhibition ELISA is very flexible, and based on this, a more complex experimental scheme can be designed to derive a special ELISA method such as direct competition inhibition ELISA, indirect competition inhibition ELISA, sandwich competition ELISA. Here, the principle is briefly explained by taking the direct competition inhibition ELISA and the indirect competition inhibition ELISA as examples. Direct competition inhibition ELISA: In a direct competitive inhibition ELISA, an antigen is pre-coated on a solid phase carrier, and an enzyme-labeled specific antibody is added. In the experiment, the antigen (or antibody) to be tested is added, and if the object to be tested is an antigen, the antigen to be tested competes with the antigen pre-coated on the solid phase carrier for the enzyme-labeled antibody; if the object to be detected is an antibody, the antibody is to be detected. The test antibody competes with the original enzyme-labeled antibody in the system to bind the antigen coated on the solid support. The washing process can wash away the enzyme-labeled antibody under competition, and finally add the substrate to develop color. The result of the final color development is inversely proportional to the amount of the original (or antibody) to be tested. Note that in the direct competition inhibition ELISA, the same pre-prepared system can measure both the antigen and the antibody. Direct competition inhibition ELISA: In a direct competitive inhibition ELISA, an antigen is pre-coated on a solid phase carrier, and an enzyme-labeled specific antibody is added. In the experiment, the antigen (or antibody) to be tested is added, and if the object to be tested is an antigen, the antigen to be tested competes with the antigen pre-coated on the solid phase carrier for the enzyme-labeled antibody; if the object to be detected is an antibody, the antibody is to be detected. The test antibody competes with the original enzyme-labeled antibody in the system to bind the antigen coated on the solid support. The washing process can wash away the enzyme-labeled antibody under competition, and finally add the substrate to develop color. The result of the final color development is inversely proportional to the amount of the original (or antibody) to be tested. Note that in the direct competition inhibition ELISA, the same pre-prepared system can measure both the antigen and the antibody. Like the indirect ELISA, the direct competitive inhibition ELISA also has a two-step signal amplification process, so its sensitivity is also relatively high, but after pre-coating the solid phase carrier and adding the enzyme-labeled antibody, the experiment only needs to dilute the antigen or antibody to be tested. It can be added to the system for reaction, which greatly simplifies the operation of the ELISA. Direct Competition Inhibition ELISA An indirect competitive inhibition ELISA can be thought of as interfering with a previously prepared indirect ELISA system with the antigen to be tested or the antibody to be detected. The specific principle is that the antigen is coated on a solid phase carrier, and a specific antibody and an enzyme-labeled secondary antibody corresponding to the antibody are sequentially added as a pre-prepared system. In the experiment, the diluted antigen (or antibody) to be tested is added, and the antigen (or antibody) in the sample to be tested competes with the antigen (or antibody) bound to the solid phase carrier in the pre-prepared system to bind to the specific antibody (or The antigen bound to the solid phase carrier is the same as the indirect ELISA. The indirect competitive inhibition ELISA also has a three-step signal amplification process, so its sensitivity is also higher than that of the direct competitive inhibition ELISA. It can be seen from the above principle that competitive inhibition ELISA requires very little reagent, and both monoclonal antibody and polyclonal antibody can be used for experiments, and more importantly, whether the subject is a macromolecular substance or a polypeptide, a small molecule drug, or small A competitive inhibition ELISA can be used for molecules such as molecular hormones that have only one epitope that cannot be used in a sandwich ELISA. In addition, in the detection of hepatitis B markers, e antigen is very unstable and easily degraded into a core antigen. Therefore, sandwich ELISA cannot be used to detect e antibodies, and only competitive inhibition ELISA can be used. In addition to the basic ELISAs described above, one can also flexibly design other types of ELISAs according to their actual needs. For example, biotin-avidin can be introduced to increase the magnification of the system, and if non-enzyme secondary antibodies are introduced, Protein A, Protein G, etc. increase the loading of the solid phase carrier or increase the specificity of the system.
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