Bio-Rad Affi-Gel 15 Gel - Manual (Page 15)

Section 8
Using the Coupled Support
When the Affi-Gel 10 or 15 activated support has
been coupled to the ligand, it is ready to use.
General operating conditions
Flow rate 15-25 cm/h
Pressure limit 15 psi
Minimum buffer concentration 50 mM
Stability
Organic solvents alcohols
pH 2-11
Temperature autoclavable (ligand permitting)
Storage 1 year at 4 °C, add 0.02% NaN
3
or
other preservative to application or
starting buffer.
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• Concentration of the ligand is too low. Protein con-
centrations of <25 mg/ml or small MW ligand con-
centrations of <15 mg/ml of gel will yield less than
optimum results.
• Ligand is too diluted. Volume of ligand should not
exceed 4.5 ml/ml of gel.
• Primary amino groups on ligand are sterically hin-
dered; add nonionic detergent (up to 1%) or a
chaotrophic agent such as 1 M guanidine HCl.
• Ligand has a molecular weight greater than 500,000.
Affi-Gel 10 and 15 supports are not suitable for this
application. Try the Affi-Prep 10 support, which is
more porous.
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