Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit - Manual (Page 29)

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c. When degassing is complete, pour the buffer into a bottle and label “Solution
A2, 50 mM Tris base”.
Solution B1
Place 1 L of water into a 1 L side-arm flask and drop in a stirbar. Cap the side
arm flask, place it on a stirplate and connect it to a vacuum source. Degas the
solution for approximately 15 minutes with gentle stirring. Label solution as
“Solution B1, water”.
Solution B2
a. Empty the contents of the bottle labeled Solution B2 into a 200 ml graduated
cylinder and add filtered, high-quality water to 125 ml. If you mistakenly
dilute solution B
2
to 500 ml, add 43.8 g NaCI, stir, and degas until dissolved.
b. Place the contents of the graduated cylinder into a 1 L side-arm flask and
drop in a stirbar. Cap the side-arm flask, place it on a stirplate, and connect it
to a vacuum source. Degas the solution for approximately 15 minutes with
gentle stirring.
c. When degassing is complete, pour the buffer into a bottle and label it as
“Solution B2, 2 M NaCl”.
4.3 Prepare Sample
a. Using the fraction collector, collect 1 ml of pH 8.1 Tris, 0% B into a tube.
Alternatively, mix 0.25 ml of Solution A1, 0.25 ml of Solution A2, and 0.5 ml
of water in an Eppendorf tube.
b. Remove the aluminum cap from the anion exchange standard vial. Slowly
remove the rubber plug from the vial (the contents may be under vacuum).
c. Add 1.0 ml of the buffer from step (a) to the vial.
d. Replace the rubber stopper and gently invert the vial to solubilize the protein
standards.
4.4 Install the UNO Q1 Column
Remove end caps from the UNO Q1 column. Keeping tubing lengths to a
minimum, connect 1/16" tubing from port 4 of the AVR7-3 inject valve to the
column inlet. Connect the column outlet to the bottom of the UV flow cell or
QuadTec flow cell. Secure the column in a vertical position.
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