3. (10 pts) Consider the available column materials (Table 3, below). Table 3. G

Question

3. (10 pts) Consider the available column materials (Table 3, below). Table 3. Gel Filtration column materials available Globular protein fractionation range (kDa S.7 Gel Type Sephadex G-10 Sephadex G-15 Sephadex G-25 Medium Sephadex G-50 Medium Sephadex G-75 Sephadex G-100 Sephadex G-200 Dry bead size (um) 40 - 120 40 - 120 50 - 150 50 - 150 40 - 120 40 - 120 40 - 120 1.5 1-5 1.5 - 30 3- 80 4 - 150 5 - 600 a. (5 pts) Which gel type would be most appropriate for separating a mixture of the following cytokines: IL-2 (15.5 kDa), IL-8 (8-9 kDa) and IL-18 (24 kDa)? Why?

Explanation / Answer

Answer: Sephadex G50 medium

Reason: Since the maximum molecular weight of the analyte is 24 kDa, it would be advisable to use a bead size which is slightly larger than it. In this way, the smallest size proteins would elute first, followed by medium size and finally the largest size in the end.

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