Chromatography Quiz (25 pts.) 1. Larger molecules elute first in gel filtration because… 2. In ion exchange chromatography, if the beads are positive, and a mixture of two proteins with opposite charges are filtered, which protein will elute first and why? 3. Reverse phase chromatography separates (large, small) molecules and is based on what type of interaction between the matrix and the proteins? ____________________ 4. Rank the following proteins in the order they will elute from a gel filtration column. Blue Dextran Myoglobin DNP-glutamate Cytochrome c Molecular Weight >500,000 16,900 313 12,400 5. Why did cytochrome c and myoglobin elute in the same fraction in experiment 1, and then separate into two fractions in experiment 2? 6. Hypothesize a possible reason for not getting fractions with good colors when you separated Mountain Dew. 7. What would have happened in experiment 3 if you had used the 35% alcohol first? 8. What does “elute” mean? 9. Why did the sugar elute in void volume in experiment 3? 10. Discuss one example of an experiment using any one of these chromatography methods to analyze results.