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The net surface charge of a protein changes with the pH. It takes the isoelectric point (pI) of this protein as the criterion. Net surface charge of a protein is zero at the pH equals to its pI. A protein carries positive net surface charge at the pH below its own pI, and it will carry negative net surface charge if the pH is raised above its pI. Researchers must choose the right method to perform protein IEX according to the net surface charge condition of the target protein at the experimental pH.