What is recommended to do to the chromatography column before use?

For the effective preparation of a chromatography column before use, soaking it in the elution buffer is essential. This practice ensures that the stationary phase inside the column has optimal conditions for binding the target molecule, such as Green Fluorescent Protein (GFP) in this context. Soaking helps to equilibrate the column, allowing it to be saturated with the solution that will be used for the purification process. This step also aids in preventing the formation of air bubbles and ensures that the column is ready to facilitate the separation of components based on their affinities.

Although vigorous shaking could seem like a way to prepare the column, it does not provide the necessary stability and equilibration for effective chromatography. On the other hand, allowing the column to sit undisturbed after it is packed may not properly condition the stationary phase for optimal performance. Additionally, storing the column in a cold environment is generally more related to maintaining the stability of the reagents or proteins, rather than the immediate preparation of the chromatography setup. Thus, soaking the column in the elution buffer is the most effective method to ensure readiness for the purification process.