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Consider a mixture of four proteins with various molecular weights. A histone molecule weighs 15 kDa, 15 kDa, a p53 molecule weighs 53 kDa, 53 kDa, an actin molecule weighs 42 kDa, 42 kDa, and an IgG molecule weighs 150 kDa. 150 kDa. Arrange the molecules in order of their elution from a gel filtration column.

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Answer:

  1. IgG
  2. p53
  3. Actin
  4. Histone

Explanation:

In gel filtration chromatography a porous matrix is used as the stationary phase. The smaller molecules are able to diffuse into this matrix while the larger ones are not. For this reason the larger molecules will elute first, followed by the smaller ones in decreasing order of size.

In other words the largest molecule will elute first and the smallest molecule will elute last:

  1. IgG
  2. p53
  3. Actin
  4. Histone

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