1. Introduction
To obtain good resolution in gel fi ltration, it is important that the column is well packed.
An air bubble or small disturbance in the medium bed will cau the sample zone to broaden. This broadening is amplifi ed as the zone migrates down the column. The result is broader peaks and lost resolution.
With traditional packing methods you often get good results. However, a disadvantage is that the medium becomes most tightly packed at the bottom of the column, instead of at the top.
GE Healthcare has developed an improved packing method which results in incread resolution. A short version of the packing method follows.
1. Inrt an adaptor at the bottom of the column.
2. Pour the medium into the column and pack the column in 2 steps.
3. Inrt the bottom piece or a cond adaptor at the top.
4. Turn the column upside-down.
The sample will be applied in the most tightly packed zone of the medium, now at
the top of the column. The result will be improved resolution.
We recommend you to follow this procedure since it has been shown to give the best results.
1.1 Equipment needed
Laboratory scale Process scale
Pump P-1, P-50, ÄKTAdesign 100 pump (P-901) ≈1800 ml/h
ÄKTAprime
system
or
Column* XK 16/40 XK 26/40 XK 50/60
XK 16/70 XK 26/70 XK 50/100
XK 16/100 XK 26/100
Cross-ctional area
cm2
of the column 2.0 cm2 5.3
cm2 19.6 Adaptor AK 16 AK 26 AK 50
Packing rervoir RK 16/26 RK 16/26 RK 50
* The fi rst number in the column name refers to the inner diameter of the column in mm.
The cond number refers to the lenght of the column i cm.
C and TricornTM columns can also be ud. The C columns must be ud with the appropriate thermostat jacket.
For preparative chromatography we recommend column XK 26/70.
Measuring cylinder, Large beaker, Buff er**, Glass rod, Small spoon or plastics spatula, (Pasteur pipette)Technical data
