What should I consider when choosing buffers for capillary electrophoresis?

There are several factors to consider when choosing buffers for capillary electrophoresis. One factor is the pKa Value. Selecting a buffer with a pKa close to the desired pH is crucial for effective buffering capacity. Buffers within the pH range of 7-9 are generally optimal for electrophoretic applications. Another factor is molecular size. Buffer ions move through the matrix without being filtered based on size, so their speed of migration depends entirely on their ratio of charge to mass. A third factor is the charges of buffer species. Buffers forming ions with high charge magnitudes can pose challenges due to increased ionic strength and conductivity. Thus, tris-borate buffer is favored for its uncharged states at the desired pH. Another factor is the solubility of a protein, which can be influenced by the surrounding pH environment. The pH of the surrounding environment can significantly impact the folding or unfolding (denaturation) of a protein. Lastly, the amount of glycosylation is another factor to consider, since it affects the function, stability, and solubility of proteins.