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Prepare the collection tubes by adding 0.1 ml of 1M Tris per ml of each fraction to be collected.
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Centrifuge or filter the sample to remove any particulates. Adjust to pH 7.0.
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Wash the column with 5 bed volumes of 20 mM Sodium Phosphate Buffer, pH 7.0.
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Apply the sample onto the column.
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Was with 5 bed volumes of 20 mM Phosphate Buffer, pH 7.0
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Elute the antibody with 1-3 bed volumes of 0.1M Glycine, collecting fractions into tubes containing 1M Tris.
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Regenerate column with 2-% Ethanol and store at 2-8oC.
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Pool fractions containing antibody and dialyze against 3 changes of PBS, at least 100 times the sample volume.
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Determine the protein concentration of the antibody. Concentrate if necessary. Store aliquots at -20oC.