What are the temperatures and methods of incubation in ELISA experiments in Hengyuan immunology?

The probe capacitor polarity needle has a head diameter of 5.8 mm. The total length of the 7-pin and 12-pin needles is 5 cm.

EL-C1600N100013-B

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During the experimental operation of the ELISA kit, incubation plays a critical role in the overall procedure. Understanding the proper temperature, methods, and precautions for incubation is essential to ensure accurate results. Here’s a detailed explanation from Hengyuan Immunology:

The incubation temperature typically ranges between 4°C, room temperature, 37°C, or even 43°C depending on the specific experiment. Among these, 37°C is the most commonly used in laboratories because it provides the optimal conditions for antigen-antibody binding. Most reactions reach their peak within 1 to 2 hours at this temperature. To speed up the process, you can slightly increase the temperature, but be cautious not to exceed the recommended limits. Additionally, shaking or oscillating the plate during incubation can enhance the contact between antigens and antibodies, leading to more efficient binding.

Incubation at 4°C allows for a more thorough reaction, resulting in more stable products. However, this method is rarely used in ELISA assays due to the extended time required. For best results, choose a temperature that balances reaction efficiency with practicality.

There are several common incubation methods, including the thermostat method, microwave irradiation, and water bath incubation. The water bath method is particularly effective in reducing the "edge effect," where the outer wells may heat unevenly compared to the center. When using a water bath, place the ELISA plate so that its bottom is in direct contact with the water surface to ensure uniform temperature distribution. To prevent evaporation, cover the plate with sealing film. If using a thermostat, place the plate inside a wet box made of a heat-conductive material like metal, with a layer of moist gauze at the bottom. Avoid stacking plates during incubation to maintain consistent temperature across all wells.