Bio-Rad Affi-Gel 15 Gel - Manual (Page 11)

3. Closed container which holds at least four times the
volume of gel.
4. (Optional) rotating shaker.
5. (Optional) Buchner funnel.
Shake the vial, and make sure that you have a uniform
suspension. Transfer the desired quantity of slurry to a
Buchner funnel or glass fritted funnel. Drain the super-
natant solvent, and wash the gel with three bed volumes of
cold 10 mM sodium acetate, pH 4.5, or cold (4 °C) deion-
ized water. The wash can be facilitated, particularly when
working with larger amounts of gel, by applying a vacu-
um. Care should be taken, however, not to allow the gel
bed to go dry. For optimum coupling of ligands, the wash-
ing procedure should be completed and the gel combined
with the ligand solution within 20 minutes.
Transfer the moist gel cake to a test tube or flask and
add the cold ligand solution. Add at least 0.5 ml of ligand
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Section 4
Recommended Storage Conditions
If Affi-Gel 10 gel is stored at -20 °C it retains at least
80% of original activity for at least 1 year. Storing at
-70 °C will extend the shelf life further.
Section 5
General Instructions
5.1 Aqueous Coupling
Material and equipment required for coupling ligands to
Affi-Gel 10 and Affi-Gel 15 gel under aqueous conditions.
1. Cold distilled water.
2. Coupling buffer of your choice without primary
amines or sulfhydryl groups; 10-100 mM HEPES,
MOPS, POPSO, acetate, or bicarbonate for proteins.
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