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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Methods Enzymol. 2014;537:243–259. doi: 10.1016/B978-0-12-411619-1.00013-6

Table 13.2.

Suggested operating parameters for FPLC operation, gel filtration, and sample collection based on a GE Superdex 200 10/300 GL column

Procedure step Flow rate (mL/min) Approx. pressure (MPa) Volume (mL) Column fractiona
Equilibration 0.6 0.6–0.7 4.8 0.2
Sample injection 0.3 0.3–0.35 0.75 0.03
Elution before collection 0.3 0.3–0.35 6.24 0.26
Fraction collection 0.3 0.3–0.35 15.6 0.65
Regeneration 0.6 0.6–0.7 21.6 0.9
Water rinse 0.6 0.6 25 1.04
Ethanol storage 0.6 0.6–0.7 25 1.04
a

Specific to Superdex 200 10/300 GL column with 24 mL packed bed.

Sample system pressures are for a column of moderate age. New columns will operate at 0.1 MPa less; older columns should be cleaned once pressures exceed 0.75 MPa. Volumes and column fractions are given to provide protocol programming guidelines following the Section 3 methodology.