Orange G and Blue dextran are in the mixture Orange G is a s

Orange G and Blue dextran are in the mixture. Orange G is a small molecule, whereas blue dextran is relatively big. Which one flows out first? Which one flows out later? Why? You need to purify a protein that is enriched with glutamate (I.e. acidic protein). What kind of chromatography would you use?

Solution

1.Blue dextran flows out first and followed by orange G

Gel filtration chromatography separates mixtures based on size. It is also called molecular exclusion or gel permeation chromatography. Small molecules enter the porous beads in the column and take longer path while the larger molecules are excluded from entering the beads in the column and elute out quicker than the small molecules.

2.Glutamate at physiological pH is a negatively charged protein, hence exists as anion. We can use anion chromatography to concentrate glutamate.

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