The Isoelectric Point (pI) is the pH at which any given protein has an equal number of positive and negative

charges, in other word the protein has no charge or neutral. At a pH below the isoelectric point, proteins carry a net positive charge, and above the Isoelectric Point protein has a net negative charge. The Implication of Isoelectric Point The property of Isoelectric Point has important biochemical implications in protein purification and electrophoresis. In electrophoresis, if the pH of the buffer is higher than the isoelectric point of the protein, it will migrate towards the positive terminal. While if the ph of the buffer is lower than the isoelectric point of the protein, then it will migrate towards negative terminal. When the buffer pH is equal to the pI of a protein it will not migrate at all. Proteins are generally least soluble at their isoelectric point, and in some cases, such as rabbit muscle glyceraldehyde-3-phosphate dehydrogenase, precipitation is achieved by adjusting the pH at constant (NH4)2SO4 concentration. But this is only useful when the isoelectric point is unusual; as mentioned, most proteins are isoelectric somewhere between 3.5 and 5. The answer is that at its isoelectric point the protein surface carries no net charge. As protein solubility is based upon favourable electrostatic interactions between the charges (negative or positive) on the protein surface and the delta-negative or delta-positive dipoles on water molecules, when there is LEAST charge on the protein surface you can also expect their to be the least favourable interactions between water molecules and the protein. As the protein-water interaction is in competition with protein-protein interactions, being at the pI most favours protein-protein interactions, which leads to precipitation, compared to protein-water interaction which lead to solvation.