Bio-Rad Bio-Scale™ Mini Affi-Prep® Protein A Cartridges User Manual
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Fig. 1. Binding capacities for murine IgG subclasses.
Mouse ascites fluids, containing
equivalent amounts (5 mg) of either mouse IgG
1
, IgG
2a
, or IgG
2b
monoclonal antibody,
were purified with 0.5 ml columns of Affi-Prep protein A matrix. Chromatography was
performed with the Affi-Prep protein A MAPS II buffers.
1.3 Kit Components
The Affi-Prep protein A MAPS II kit contains enough reagents to perform over 100
analytical cartridge runs, for purification of up to 500 mg of murine IgG.
Affi-Prep protein A: One 30 x 4.6 mm Affi-Prep protein A analytical cartridge (0.5 ml
bed volume; catalog number 125-0460) in 0.1 M HEPES, 1 mM
EDTA, pH 7.5, containing 0.05% sodium azide.
Cartridge holder:
One Standard Cartridge Holder (catalog number 125-0131).
Binding buffer:
One bottle (471 g) of buffer solids. Reconstituted final volume =
1,500 ml.
Elution buffer:
One bottle (25 g) of buffer solids. Reconstituted final volume =
1,100 ml.
1.4 Additional Items Required
pH meter:
A pH meter is required to check the pH of binding and elution
buffers after reconstitution.
Mixer:
Standard laboratory magnetic stirrer and stir-bar for buffer mixing.
Balance:
Standard laboratory scale for weighing of buffer solids.
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