What should be predicted when adding Wash Buffer to the column?

When adding Wash Buffer to the column during the purification process of Green Fluorescent Protein (GFP), the anticipated outcome is that non-target proteins may wash off. The Wash Buffer is designed to remove proteins that are not specifically bound to the column's stationary phase, allowing for the selective elution of desired proteins. This process relies on the differences in binding affinity between target proteins, like GFP, and non-target proteins.

As the Wash Buffer passes through the column, it helps to dislodge those proteins that do not interact strongly with the column matrix, resulting in their removal. This step is crucial in enriching the desired protein, as it minimizes contamination from other cellular proteins. The specific conditions of the Wash Buffer, such as its ionic strength or pH, are optimized to ensure that only the weaker-bound, non-target proteins are eluted, thereby enhancing the purity of the GFP that remains bound to the column.

The other outcomes, like the elution of all proteins or the stickiness of hydrophobic proteins to the column, do not accurately reflect the purpose of adding Wash Buffer in this context. The primary objective is to selectively remove impurities while retaining the target protein, which is effectively achieved by understanding how different proteins interact with the column media.