Bio-Rad Media Sampler Pack - Manual (Page 5)

Table 1. Characteristics of Macro-Prep HIC Media
Macro-Prep t-Butyl Macro-Prep Methyl
Functional groups COO- and t-butyl COO- and methyl
Ionic capacity Approximately 120 µeq/ml Approximately 120 µeq/ml
Shipping counterion Cl- Cl-
Hydrophobicity Mild Mild
Median particle size (µm) 50 50
pH stability 1–10 1–10
Autoclavability (121°C, 30 min) Yes Yes
Shipping solution 20% ethanol 20% ethanol
Working flow-rate range 100–600 cm/hr 100–600 cm/hr
Sanitization 5 CV of 6 M guanidine-HCl, 5 CV of 6 M guanidine-HCl,
100 cm/hr 100 cm/hr
or or
5 CV of of 1% acetic acid 5 CV of 1% acetic acid
in 0.12 M phosphoric acid, in 0.12 M phosphoric acid,
pH 1.5, 100 cm/hr pH 1.5, 100 cm/hr
Storage at 4–40°C 20% ethanol 20% ethanol
or or
1% acetic acid in 0.12 M 1% acetic acid in 0.12 M
phosphoric acid, pH 1.5 phosphoric acid, pH 1.5
Section 5
Preparation for Use
Macro-Prep HIC media are supplied fully hydrated in 20% ethanol as a 50% slurry.
Small volumes of Macro-Prep HIC media are easily washed in a Büchner funnel
with 4–5 column volumes (CV) of a low ionic strength buffer (<20 mM). For large
process-scale volumes, pack the column in a low ionic strength buffer (<20 mM)
or water and then rinse with at least 3 CV of the low ionic strength buffer or water
before transferring to the high-salt buffer. Alternatively, decant the ethanol shipping
solution and resuspend/decant 3–4 times in the low ionic strength buffer to be
used for column packing.
Section 6
Column Packing
Columns can be packed in different ways depending on the media and type of
column and equipment to be used. Macro-Prep HIC media can be packed using
pressure, flow, or suction packing methods. To pack highly efficient columns, it is
recommended to use a 50% slurry. Some general guidelines for packing small and
large columns are given below. Make sure to read and follow the instructions for
the column to be used. If unsure about a particular column, call your local Bio-Rad
Laboratories representative. Throughout this instruction, flow rate is expressed
linearly in centimeters per hour (cm/hr). The relationship between linear and
volumetric flow rate as delivered by a pump is:
Linear flow rate (cm/hr) X Cross-sectional area of the column (cm 2 ) = Volumetric flow rate
Example: 300 cm/hr in a 14 cm diameter column = 300 cm/hr X πr 2 or 300 cm/hr X
(3.14 X 7 2 ) cm 2 or 46,158 cm 3 /hr or 769.3 ml/min.
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