Discussion
Which of the following technique(s) can be used to separate proteins according to their molecular weights from a mixture of proteins?
Answer: B, C, D
Options B, C and D are correct; the key accepts all three. D is the weakest, since sedimentation also depends on shape.
Size exclusion chromatography separates by hydrodynamic size: large molecules elute first. SDS-PAGE coats proteins with a uniform negative charge, so migration depends on size and gives molecular weight. Sucrose density gradient centrifugation sediments larger proteins and complexes faster, so it separates by mass, with the caveat of shape.
Ion exchange chromatography (A) separates by net charge at the working pH, not by molecular weight.
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