During the experimental operation of the elisa kit: Incubation is an important step in the experimental procedure. What are the temperatures and methods of incubation and the precautions? Hengyuan Immunology explains to you:
Incubation temperature: The temperature used for breeding is 43 °, 37 ° C, room temperature or 4 ° C. 37 ° C is the incubation temperature commonly used in the laboratory, and is also the optimum reaction temperature for most antigen-antibody binding. The antigen-antibody reaction generally reaches a peak at 37 ° C for 1 to 2 h. To accelerate the reaction, the reaction temperature can be increased within a certain range and continuously oscillated during the reaction to increase the probability of antigen-antibody contact. The antigen-antibody reaction is more thorough at 4 ° C, and the resulting product is more and more stable, but it is generally not used in ELISA assays because the time required is too long.
The way of incubation: the incubation method often uses the thermostat method, the microwave irradiation method and the water bath method. Among them, the water bath method can better solve the difference in absorbance (ie, "edge effect") between the surrounding pores and the central pore due to uneven heat. The ELISA plate can be placed in a water bath, and the bottom of the plate should be placed against the water surface to quickly balance the temperature. To avoid evaporation, the plate hole can be covered with plastic sealing paper. If a thermostat is used, the ELISA plate 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. Each reaction plate cannot be placed in an overlapping process during the incubation process to prevent unevenness in temperature diffusion.
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