Bio-Rad Bio-Scale™ Mini CHT™ 40 µm Cartridges - Manual (Page 11)

Table 3. Products for Buffer Exchange
Sample Recommended
Volume
Product Use Catalog #
50–100 µl Bio-Spin 6 column Desalting proteins 732-6000
≥6 kD
50–100 µl Bio-Spin 30 column Desalting proteins 732-6004
≥30 kD
100 µl–3 ml Bio-Scale Mini P6 Desalting proteins 732-4502
cartridge ≥6 kD
Up to 3 ml Econo-Pac 10DG Desalting proteins 732-2010
desalting columns ≥6 kD
Unlimited Bio-Gel P-6DG gel Desalting proteins 150-0738
≥6 kD
4.2 General Purification Protocol
Hydroxyapatite chromatography is usually performed
using increasing linear salt gradients to elute the
sample components. For best results, and increased
cartridge life, samples and buffers should be degassed
and filtered through a 0.45 µm filter. In order to
promote the stability of CHT, buffers are recommended
to include low concentrations of either phosphate or
15
5. Equilibrate the cartridge with running buffer and
operatate the cartridge according to the CHT
instruction manual or according to your protocol.
4.1 Sample Preparation
Proper 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's concentration. This can be
achieved by diluting the sample to the ionic strength
of the starting buffer, dialyzing against the starting
buffer, or exchanging it into the starting buffer.
Buffer exchange can be accomplished using the
Bio-Scale Mini P6 cartridge, Bio-Spin
® 6 or
Bio-Spin 30 columns, Econo-Pac 10DG desalting
columns, or Bio-Gel
® P-6DG gel filtration gel. The
choice of product will depend on sample volume.
All samples should be filtered through a 0.45 µm
filter prior to cartridge application.
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