(1) Draw a chromatogram depicting separation of proteins 1, 2, and 3 (protein #1, mw 30,000, protein #2, mw 60,000, and protein #3, mw 90,000). Which type of packing material would you use to separate these proteins?
(2) Indicate on the chromatogram in (1) where insulin would most likely elute. Would it have total accessibility, some accessibility or no accessibility to the particle pores on this column?
(3) Referring to the size exclusion chromatogram in (1), Indicate on the chromatogram where you would expect a molecule with a size of 40,000 to elute. Would you expect the molecule to be well resolved, somewhat resolved, or not resolved from the peak for protein #1? Explain.
(4) Use the SEC calibration plot shown below to estimate the molecular weight (MW) of an unknown protein with a retention volume of 3mL.