Bio-Rad Profinia Protein Purification System Kits - Manual (Page 15)

conical tube and place on ice until ready to start the purification. The
remaining 6 ml of resuspended sample can be frozen for later use and
thawed once prior to purification. The frozen sample should be used within
6 months.
Remove the bottle of GST elution buffer and vial of reduced glutathione.
Transfer 10 ml elution buffer to the vial of glutathione, shake to mix, and
let stand 5 min at room temperature to completely dissolve. Transfer the
resuspended glutathione to the 125 ml bottle of elution buffer. The elution
buffer (2x) now contains 40 mM glutathione, which is necessary to elute
GST-tagged proteins. The remaining bottles of GST buffers and cartridges
require no advance preparation.
5.2 Instrument Startup, User Interface, and Method
Selection
Turn on the instrument using the recessed on/off switch at the right
lower panel. Make sure the cleaning tray is in place and that filtered water
is in buffer positions 1–7. Choose Data/Utilities, Diag/Maint Functions, and
Clean All Inlet and Outlet Lines. This will flush ethanol or air from the system
prior to starting the chromatography run. After the system is rinsed press
Home to return to the Profinia home screen. Choose Bio-Rad Methods.
The user interface for the GST starter kit is similar to that described in
Section 12. Choose GST on the select method screen. The method
screen for the GST method type and options should look like Figure 11.
Select Method Type and Options
Screen
The Select Method Type and Options
screen allows the user to quickly
choose the method of purification,
number of samples to be purified, and
the column volume (ml) for the affinity
and/or desalting cartridges. The
selections in Figure 11 are:
• Method type: GST + Desalting
• Sample type: 1 sample
• Cartridge selection: 1 cartridge
11
Fig. 11. Method selection for GST application.
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