Two monomeric His-tagged proteins of identical molecular weight are present in a solution. Their pI values are 5.6 and 6.8. Which technique can separate them?
The proteins have the same size, so size exclusion and ordinary denaturing PAGE do not exploit the specified distinction. Both carry His tags and bind nickel affinity media. Their different pI values produce different net charges at a selected pH; ion exchange separates proteins on that basis. For example, between the two pI values their net charges have opposite signs. Select C.
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