Bio-Rad Bio-Scale Mini Affi-Gel Blue Cartridges User Manual
Bio-Scale™Mini Affi-Gel
®
Blue and DEAE Affi-Gel
Blue Cartridges, 5 ml
Instruction Manual
Catalog #
732-4642, 732-4644
732-4632, 732-4634
Table of Contents
Section 1...Introduction ........................................1
Section 2 Connecting to Bio-Rad’s
Low-Pressure Chromatography
Instruments ........................................5
Section 3 Connection to Other Liquid
Chromatography Systems................10
3.1 Connecting to BioLogic
DuoFlow
™
Systems ...............10
3.2 HPLC Systems ......................11
3.3 FPLC Systems .......................12
Section 4 Preparing a Cartridge for Use...........13
4.1 Sample Preparation ...............16
4.2 General Purification
Protocol .................................18
4.3 Scaling Up the Separation .....18
Section 5 Specific Purification Protocols ..........19
5.1 Purification of IgG from
Serum or Ascites with
the Bio-Scale Mini DEAE Affi-Gel
Blue Cartridge........................19
5.2 Purification of Serum
Proteins with the
Bio-Scale Mini DEAE Affi-Gel
Blue Cartridge........................20
5.3 Removal of Albumin from
Serum with the Bio-Scale Mini
Affi-Gel Blue Cartridge ...........21
5.4 Purification of Enzymes
with the Bio-Scale Mini Blue
Affi-Gel Cartridge ...................22
Section 6...Care of the Cartridge........................23
6.1 Cleaning the Cartridge ...........23
6.2 Storage..................................24
Section 7 Technical Assistance ........................25
Section 8 Ordering Information.........................26
Section 9 References .......................................28
Section 1
Introduction
Bio-Scale Mini cartridges have a patent-pending
double-wall design that provides extra durability and
allows easy, reliable runs with aqueous buffers
commonly used for protein purification. The
polypropylene luer fittings and internal sealing
surfaces ensure leak-free operation at pressures up
to 45 psi. Bio-Scale Mini cartridges are convenient,
disposable, and supplied ready for use. They are
easy to use and prepacked for fast, reproducible
chromatographic separations. Cartridges are
available for a variety of chromatographic techniques,
including desalting, ion exchange, affinity, and
mixed-mode chromatography. The design of
Bio-Scale Mini cartridges offers:
• Ready-to-use convenience; simply equilibrate
the cartridge in the buffer of choice
• Luer fittings for convenient connection to any
chromatography system or directly to a Luer-Lok
syringe
1
Bio-Scale Mini Affi-Gel Blue and DEAE Affi-Gel Blue
cartridges are packed with Affi-Gel
®
affinity
chromatography gels. These gels are hydrophilic
spherical crosslinked agarose beads designed for
the purification of proteins or the removal of
high-abundance proteins from cell lysates or serum.
Request bulletin 1107 for other applications. Affi-Gel
beads are designed to provide medium capacity,
low backpressure, and high productivity. Detailed
product information is given in Tables 1 and 2. See
Ordering Information for a listing of the complete
Bio-Scale Mini cartridge product line.
2
Table 1. Bio-Scale Mini Affi-Gel Blue and
DEAE Affi-Gel Blue cartridge specifications.
Size 5 ml bed volume
Dimensions 40 mm length x 12.6 mm inner diameter
Maximum pressure tolerance 10 psi (680 mbar. 0.36 mPa)*
Operational flow rates 0.5–2.0 ml/min (25–100 cm/hr)
Fittings: Female luer fitting inlet;
male luer fitting outlet
Column material Polypropylene
Frit material Polyethylene (HDPE)
Shipping conditions 0.02 M sodium phosphate, pH 7.4,
0.05% NaN
3
Storage recommendations 0.02 M sodium phosphate, pH 7.4,
0.05% NaN
3
Autoclavability Not autoclavable
*Although the pressure limitation of the cartridge is 45 psi, the agarose gel
tends to compress above 10 psi (generally 3.0 ml/min).
3
Table 2. Affi-Gel Blue and DEAE Affi-Gel
Blue specifications.
AAffffii--GGeell BBlluuee DDEEAAEE AAffffii--GGeell BBlluuee
Type Dye affinity Dye affinity/weak anion
Functional group Cibracron Blue F3GA Cibracron Blue F3GA
and – N(C
2H5)2
Serum capacity 0.3–1.0 ml 0.3–1.0 ml
Maximum flow rate 3.0 ml/min 3.0 ml/min
Operating pH range 2–10 2–10
pH stability 1–10 1–12
Chemical compatibility See Table 3.
4
Section 2
Connecting to Bio-Rad’s
Low-Pressure
Chromatography Instruments
Bio-Scale Mini cartridges are ideal for use with
Bio-Rad's BioLogic
™
LP chromatography system,
Econo
™
gradient pump, the patented* Model EP-1
Econo
™
pump, and all low-pressure chromatography instruments. Bio-Scale Mini cartridges can be
conveniently connected directly to the system using
the luer fittings on the cartridge.
1. Install 1.6 mm inner diameter (ID) tubing in the
pumphead. Adjust the platen pressure screw (on
pumphead) – using a screwdriver or coin, turn
the screw counterclockwise as far as it will go,
then turn clockwise three full turns. Assemble
with fittings and lock rings as shown in Figure 1.
*US Patent 5,135,658
5
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