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Two monomeric His-tagged proteins of identical molecular weight are present in a solution. pIs of these two proteins are $5.6$ and $6.8$. Which one of the following techniques can be used to separate them?

  1. Denaturing polyacrylamide gel electrophoresis
  2. Size-exclusion chromatography
  3. Ion-exchange chromatography
  4. Nickel affinity chromatography
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