Why do pig ELISA kits need insulation?

Why does the pig ELISA kit need to be kept warm? Because the ELISA is a solid phase immunoassay, the binding of antigen and antibody occurs only on the surface of the solid phase. Taking the sandwich method of antibody coating as an example, the specimen added to the well of the plate does not have an equal opportunity to bind to the solid phase, and only the antigen in the solution closest to the pore wall is directly in contact with the antibody. . This is a process of gradual equilibrium, so it needs to be diffused to reach the end of the reaction.

There are generally two antigen-antibody reactions in an ELISA, ie, after adding the sample and adding the enzyme conjugate. Insulation method In addition to the special ELISA instrument with special electric heating block, the water bath is generally used. The ELISA plate can be placed in the water bath box. The bottom of the ELISA plate should be attached to the water surface to make the temperature balance quickly. In order to avoid evaporation, the board should be covered, or the plate hole can be covered with plastic sealing paper or plastic wrap. At this time, the reaction plate can float on the water surface.

If an incubator is used, the ELISA kit should be placed in a wet box. The wet box should be made of a material with good heat transfer such as metal, a wet gauze at the bottom of the box, and finally the ELISA plate should be placed on the wet gauze. The wet box should be pre-warmed in the incubator to the specified temperature, especially at lower temperatures. Whether it is water bath or wet box incubation, the reaction plates should not be stacked to ensure that the temperature of each plate can be quickly balanced. For the reaction at room temperature, the room temperature during the operation should be strictly limited to the specified range. The standard room temperature refers to 20 to 25 ° C, but the specific conditions can be controlled according to the requirements of the instructions.

When incubated at room temperature, the pig ELISA kit can be placed flat on the console. It should be noted that the temperature and time of incubation should be as accurate as required. In order to ensure this, one person should not measure more than two plates at the same time.

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