Bio-Rad Foresight™ Chromatography Columns, Prepacked - Manual (Page 10)

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Section 3
Preparing a Column for Use
Foresight™ columns may be shipped to you containing 20%
ethanol, 20% ethanol + 0.15 M NaCl (v/v), or dry, depending on
the chromatography media. All columns are ready to use after
equilibrating the columns in the buffer of choice. To perform a
buffer exchange, connect the column to a liquid chromatography
system or peristaltic pump and condition it as follows:
1. Set the pump flow rate to 0.5–1 ml/min (59.5–119 cm/hr) for
the 1 ml column or 2.5–5 ml/min (297.5–595 cm/hr) for the
5 ml column.
2. Wash the column with degassed low-salt buffer for 2
column volumes. Wash with degassed high-salt buffer for 5
column volumes.
3. Equilibrate with low-salt buffer for 5 column volumes.
4. Reduce the flow rate to the rate that will be used in the
purification protocol.
3.1 Sample Preparation
Correct pH and ionic strength are necessary for consistent
and reproducible results. Sample can be exchanged
into the starting buffer or diluted to the starting buffer
concentration. This can be achieved by diluting the sample
to the ionic strength of the starting buffer, dialyzing against
the starting buffer, or exchanging into the starting buffer.
Buffer exchange can be accomplished using a number of
products (Table 3). The choice of product will depend on
sample volume. Filter all samples through a 0.45 µm filter
prior to column application.