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The rabbit sera Wash the column with 40 ml PBS (if antibody shows unspecific binding, washes. Affinity Purification of Antibodies. Day 1: Coupling of ligands to beads. Ligand amounts for coupling: protein 5-10 mg per ml beads peptide 1- 5 mg per ml beads. Affinity Purification Please note that this protocol is designed for affinity purifying additional serum using the column that we have created for you. To make an antibody affinity purification column, minimally 3mg of protein or coupled peptide is required, but more is definitely desirable.

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Try and aim for about The following is a general protocol for affinity purification of antibodies from serum. Peptides or proteins are covalently bound to agarose beads by their free Common elution buffer systems for protein affinity purification. These conditions apply primarily to protein-protein binding interactions, such as between an antibody and its peptide antigen.

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