Pouring slurry into the column
This section assumes that the column has been primed.
1
Resuspend the slurry in the container of medium by gentle shaking until fully homo-
geneous.
Ensure that slurry concentration matches that entered into the wizard.
Note:
2
Pour the appropriate volume of resuspended slurry (see Column Packing Summary
in Intelligent Packing wizard or in the method notes) into the column tube.
It is preferable to pour the gel slurry against the inner wall of the column
tube to minimize air trapped in the slurry. Do not however tilt the column
in order to achieve this.
Note:
3
Wash out any remaining medium from the container and pour it into the column.
4
Use the media stirrer to gently agitate the medium in the slurry so that there are no
clumps of medium and that the slurry is well mixed. Make sure that there is no sedi-
mented medium left at the bottom of the column.
It may be necessary to lower the level of the slurry in the column tube to
accommodate volume displacement by insertion of the media stirrer and
additional liquid volume from rinsing.
Note:
5
Add more packing liquid into the column right up to the edge of the glass column
tube. Medium stuck on the column wall or in the hydraulic chamber inlet should be
flushed down into the gel slurry with packing liquid. Adding too little liquid increases
the risk for air bubbles becoming trapped, whereas too much liquid will lead to
spillages when the adapter is inserted.
6
Let the column stand undisturbed so that the medium can begin to settle. The speed
of settling depends on the medium. Wait until there is a clear head of liquid 1-2 cm
above the level of the settling medium before inserting the adapter. This reduces the
risk of medium being forced up beyond the lower surface of the adapter. Do not wait
more than 15 minutes to avoid a size gradient in the bed.
NOTICE
Insert the adapter into the mouth of the column tube at an
angle to ensure that no air is trapped in the column.
7
Fit the adapter and top plate onto the column (see Fitting the top plate and adapter,
on page 41).
74
AxiChrom 50, 70 and 100 columns Operating instructions 28-9331-08 AG
5 Operation
5.1 Intelligent Packing of the column with ÄKTAexplorer and ÄKTApilot
