Bio-Rad ENrich™ High-Resolution Size Exclusion Columns - Manual (Page 7)

ENrich SEC Instruction Manual 3
Section 2 : Use of the ENrich
Columns
2.1 Preparation for Initial Use
The column is supplied in 20% ethanol. To remove the storage
buffer, pump deionized, filtered water at a flow rate of 0.5 ml/min
or less for 30 ml, or just over one column volume (CV). The
backpressure will decrease as the ethanol is washed from the
column. After this water step you may equilibrate your column as
described below.
Warning: Do not exceed a maximum pressure of 600 psi
(4.1 MPa, 41 bar).
2.2 Equilibrating and Running the Column
1. Always use filtered (0.22–0.45 μm filter) and degassed buffers.
This will prolong the life of your column. To avoid bacterial
growth, contamination, and poor column performance, use
only freshly prepared buffers.
2. Residual ionic charges on the ENrich column are negligible.
However, a running buffer with ionic strength of at least
100 mM NaCl is recommended. Phosphate buffered saline
(PBS) is an example of a commonly used buffer appropriate for
ENrich SEC media.
3. The buffer should contain any cofactors or protease inhibitors
previously identified as being essential to maintain enzyme
activity.
4. Equilibrate the column with at least 2 CV (~50 ml) of the buffer.
5. Calibrating the column with a gel filtration standard
(catalog #151-1901) is an important step for molecular weight
determination and for tracking column performance.