Bio-Rad Macro-Prep 25 S Media - Manual (Page 10)

8
Cleaning-in-Place (CIP)
Acceptable CIP agents include 0.4 M NaCl in 1% acetic acid/1% phosphoric acid,
up to 30% acetic acid, 1% Triton X-100, up to 70% ethanol or up to 30%
isopropyl alcohol, 8 M urea, and 6 M guanidine-HCl. Any of these agents can be
combined in an appropriate cleaning protocol, which is often developed empirical-
ly. As a general guide, we recommend the following:
1. Use high salt buffer for regeneration, as above.
2. Remove additional contaminants with 0.4 M NaCl in 1% acetic acid/1%
phosphoric acid (3–5 CV) at 100 cm/hr.
3. For aggregated or precipitated proteins, or when dirty feedstock (for example,
crude lysate) has been used, wash with 3–5 CV of 6 M guanidine-HCl or
8 M urea at 100 cm/hr.
4. For lipids or hydrophobically bound contaminants, wash with 0.1% Triton
X-100, or 20–70% ethanol or 20–30% isopropyl alcohol, or 10–30% acetic
acid. Use 3–5 CV at 100 cm/hr.
5. Finally, wash with 2 column volumes of deionized water and 4–5 CV of
starting buffer. Check the conductivity and pH of the effluent to verify that the
column is equilibrated in the starting buffer before loading the sample.
Sanitization and Storage
To sanitize and store between campaigns, wash the column with 3–5 CV of 1%
acetic acid/1% phosphoric acid, pH 1.5. Store the column at 4–40°C. When not in
use, store the Macro-Prep 25 ion exchange supports in either 1% acetic acid/1%
phosphoric acid (pH 1.5) or in 20% (v/v) ethanol solution or in 2% benzyl alcohol.
The Macro-Prep 25 ion exchange supports may also be autoclaved at 121°C,
2 bar, in a neutral pH slurry, for up to 30 min and stored in one of the above
solutions.
Shelf Life
The Macro-Prep ion exchange supports are stable for at least one year when
stored sealed in the original container at room temperature.
4006129B.qxd 6/13/2005 1:55 PM Page 8