Size exclusion chromatography:
1. list the three types of column chromatography and explain how each technique separates molecules.
2. briefly explain how the chromatography used in this lab functions.
3. if the following mix of molecules were purified using size exclusions chromatography, what would be the order in which the molecules pass through the opening in the bottom of the column? mixture containing: hemoglobin 65,000 daltons; myoglobin, 17,000 daltons; myosin, 180,000 daltons.
first molecule to appear:
second molecule to appear:
third molecule to appear:
4. if a size exclusion chromatography column is said to have an exclusion limit of 40,000 daltons, would hemoglobin (65,000 daltons) be fractionated or excluded from the column?
5. would vitamin b12 (1,350 daltons) be fractionated or excluded from the column?
6. why did you need to add more buffer after the protein mixture was loaded onto the column?