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The globular protein ferritin, isolated from horse spleen, has the molecular weight of about 440,000 daltons....

The globular protein ferritin, isolated from horse spleen, has the molecular weight of about 440,000 daltons. If we had added it to the sample, what would its elution volume be? Why?

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Answer #1

Gel filtration chromatography is used to determine the molecular weight of proteins. In this case molecules are passed from gel matrix and measured against standard. The standard have a definite known value and we can estimate molecular weight by plotting the graph.

In our case no standard or any other value of protein is given. However the molecular weight of ferritin is 4,40,000 Da, which is very high. “the elution volume of high molecular weight protein is same as the column void (how much the column is empty) volume”. Hence volume of elution is same as void volume of our column. Because larger or high molecular weight protein have low mobility in gel matrix. The amount of protein is passes from gel matrix which creates the empty space in column is equal to elute.

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