The characteristics and operating recommendations for individual media are described
in instructions included with each container of medium.
Slurry concentration
To measure the slurry concentration:
1
Gently shake the container of medium until homogeneous.
2
Quickly remove the lid of the container and pour the contents into a graduated
cylinder. Let the medium settle (overnight for approximate bead size of 90 µm, and
48 h for < 50 µm) so that you can read the volume ratio of sedimented medium. The
cylinder should be placed on a flat surface and covered with for example Parafilm,
to minimize evaporation.
Some media may need more than 48 h to settle completely.
Note:
Once the settled bed volume has been obtained, the correct volume of slurry can
be calculated based on the slurry concentration. The slurry concentration in new
containers of media is approximately 70% (v/v).
3
Before packing, replace the storage liquid with a suitable packing liquid. For example:
a
Decant the storage liquid from the top of the settled medium, add new liquid
and gently mix. Repeat the decanting step several times, or;
b
Pack the column with medium, wash with several column volumes of appropriate
liquid, and repack the column, or;
c
Filter smaller volumes of medium using a vacuum filtration system with a glass
filter funnel.
Slurry volume
The method wizard calculates and presents the needed slurry volume in the wizard
summary page according to the following formula;
Slurry volume =
Target bed height x PF x Column cross sectional area x SVF
Slurry concentration
where the Slurry Volume Factor (SVF) is a compensation factor for the difference between
a gravity settled bed height and a consolidated bed height, and PF is the Packing Factor,
(see Definitions associated with the Packing Factor, on page 18).
The SVF is valid for slurry concentration measurements performed with media in 20%
ethanol and beds packed with the packing liquid suggested in the wizard summary page.
If the packing liquid used is different from the packing liquid recommended in the wizard
summary page, or if a custom medium choice is made, the SVF needs to be estimated
by the user. This is done by repeated comparisons of the gravity settled bed in a cylinder
with the achieved packed bed height in the preferred packing liquid.
All S and SP ligand media are shipped and supplied in 0.2 M sodium acetate
and 20% ethanol. Any decanted solution will contain salt.
Note:
64
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
