Table 1. UNO Q Column Characteristics
Q-1 Q-6 Q-12
Column volume (ml) 1.3 6 12
Recommended max. protein loading (mg) 20 80 160
Recommended flow-rates (ml/min) 0.5 to 5.0 0.5 to 8.0 0.5 to 10.0
Column Dimensions (mm) 7 x 35 12 x 53 15 x 68
Maximum operating pressure (psi/MPa/bar) 700/4.5/48 700/4.5/48 700/4.5/48
Table 2. UNO S Column Characteristics
S-1 S-6 S-12
Column volume (ml) 1.3 6 12
Recommended max. protein loading (mg) 20 80 160
Recommended flow-rates (ml/min) 0.5 to 6.0 0.5 to 8.0 0.5 to 10.0
Column Dimensions (mm) 7 x 35 12 x 53 15 x 68
Maximum operating pressure (psi/MPa/bar) 700/4.5/48 700/4.5/48 700/4.5/48
Column Hygiene
Careful preparation (especially filtration) of the sample and the buffers will maintain the
column performance and lifetime. Normally, washing with 1.0 M NaCl or KCl will remove
most bound components. However, if there is a significant decrease in column performance
(i.e. increasing backpressures or a significant drop in resolution) then a more extensive clean-
ing protocol as described below (steps 1–6) should be used. Always reverse the flow during
this procedure so tightly-bound substances at the top of the column are quickly removed.
This process should be performed within the following flow rates.
S-1 or Q-1 1 ml/min
S-6 or Q-6 2 ml/min
S-12 or Q-12 3 ml/min
1. Wash with 2 column volumes of 2.0 M NaCl or KCl.
2. Wash with 1 column volume of 2.0 M NaOH followed by 3 column volumes of 0.1 M
NaCl.
3. Wash with 1 column volume of 50% acetic acid followed by 3 column volumes of 0.1 M
NaCl. Wash with 1 column volume of deionized water.
4. If lipid contamination is a problem, wash with 1 column volume of MeOH followed by
1 column volume of deionized water and 3 column volumes of 0.1 M NaCl.
5. Wash with 2 column volumes of 2.0 M NaCl or KCl or the salt containing the desired
counter-ion.
6. Change the top frit and wash with 3 column volumes of start buffer.
References
1. Kopaciewicz, W., Rounds, M. A., Fausnaugh, J. and Regnier, F. E. Retention Model for High-per-
formance Ion-Exchange Chromatography, J. Chromatography, 266, 3-21 (1983).
2. A Guide to the Properties and Uses of Detergents in Biology and Biochemistry. J. Neugebauer
(Calbiochem Biochemicals).
3
(28 Seiten)
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