Mini Program in Chromatography (B3 1-3)
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1. Separating Molecules by Gel Filtration
Students fill their chromatographic columns with packing material and separate three colored molecules of known molecular weights as shown in the photograph below. The exercise illustrates how components of a complex mixture can be separated by chromatography and introduces the concept of the standard curve for determining the size of an unknown protein.
2. The Molecular Weight of Hemoglobin
Students determine the size of hemoglobin and estimate the number of amino acid residues in this protein.
3. Binding Specificity of Serum Albumin
The binding of an enzyme to its substrate is only one example of the many specific molecular interactions that occur in biological systems. An analogous binding process occurs with serum albumin which binds certain small molecular weight molecules in blood. In this exercise, students use a gel filtration assay to examine the binding of various dyes to albumin and determine if the native shape of the protein is required for its binding specificity.
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