GE AKTApure, User Manual, Page: 153 / 522

CAUTION
Risk of explosion. Do not use Mixer chamber 15 ml with an ÄKTA pure 25 configura-
tion. The maximum pressure for Mixer chamber 15 ml is 5 MPa.
In ÄKTA pure 25 at low flow rates (below 0.2 ml/min) a gradient of sufficient quality may be
achieved with the mixer bypassed. Similarly, for low flow rates in ÄKTA pure 150 the 0.6 ml
mixer might be used.
Note:
The 1.4 ml mixer in ÄKTA pure 25 or 5 ml mixer in ÄKTA pure 150 might work up to maximum
flow rate provided that the buffers are easily mixed. Note:
For information on how to install a Mixer chamber, refer to Section 7.8.3 Replace the Mixer, on page 322.
Select UV flow cells
General considerations
Flow cells with shorter path lengths are suitable to use when high protein concentrations are used.
Flow cells with longer path lengths are suitable to use when low protein concentrations are used.
UV flow cells for UV monitor U9-M
UV flow cells are available with three different path lengths; 0.5 mm, 2 mm (default) and 10 mm.
The real cell path length of the UV cell is automatically recognized by the monitor when a cell is fitted.
The UV data is normalized to the nominal path length. This allows UV data from runs made with different
UV flow cells (but with the same nominal path length) to be directly compared.
UV flow cells for UV monitor U9-L
UV flow cells are available with two different path lengths; 2 mm (default) and 5 mm. When replacing
a UV flow cell, the path length must be set in the System Control module, in System:Calibrate. Use
the nominal flow cell length if the UV flow cell is replaced but not calibrated. See Update the cell path
length , on page 314 to set the flow cell path length.
The path length of the UV flow cell might differ from the nominal length, which leads to incorrect results
in the calculation of protein concentration in the eluate. The UV data is normalized to the nominal path
length. This allows UV data from runs made with different UV flow cells (but with the same nominal
path length) to be directly compared. To achieve normalized absorbance, the path length of the UV
flow cell must be calibrated and the calculated flow cell path length set manually. See Calibration of
the UV monitor U9-L flow cell length, on page 312.
Prepare the inlet tubing
Connect inlet tubing to the inlet ports that are to be used, and place all inlet tubing that is to be used
during the method run in the correct buffers.
When using high viscosity buffers/samples in combination with high flow rates it is recom-
mended to increase the tubing I.D. and/or shorten the length.
Note:
ÄKTA pure User Manual 29119969 AB 153
5 Operation
5.2 Prepare the flow path