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Protein G Purification of Antibodies
(Protocol summary only for purposes of this preview site)IgGs from most species bind to Protein G at near physiological pH and ionic strength with a higher affinity than IgG binding to Protein A. Therefore, the pH required to dissociate bound IgG is lower, resulting in the loss of activity for some antibodies. If this is observed, Protein A may be an alternative (Protocol 5) if the IgG from the species being isolated can be purified using Protein A. Typically the binding capacity of Protein G is 1025 mg of human IgG or 510 mg of mouse IgG per milliliter of resin. Protein G covalently attached to cross-linked agarose or other matrices is available from many different vendors.
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