Thermo Scientific Pierce Protein A/G Plus Agarose is an exceptionally high-capacity Protein A/G beaded agarose resin for use in a variety of antibody affinity purification methods.
Pierce Protein A/G Plus Agarose consists of purified Protein A/G recombinant fusion protein that has been covalently immobilized at high density onto high-quality crosslinked 6% beaded agarose (CL-6B). Because the Protein A/G combines the immunoglobulin-binding domains of both Protein A and Protein G, the resin is effective for affinity purification of IgG antibodies from a broad range mammalian host species.
- Protein A/G – immobilized recombinant fusion protein of the antibody-binding domains of Protein A and Protein G enables polyclonal IgG purification from nearly any mammalian species
- Agarose resin – support is crosslinked 6% beaded agarose (CL-6B), the most popular resin for protein affinity purification methods
- Inert and stable – superior manufacturing method immobilizes Protein A/G by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
- High capacity – this “Plus” variety of Pierce Protein A/G Agarose has a very dense load of immobilized Protein A/G, providing a binding capacity greater than 50mg human IgG/mL resin
Protein A/G is a genetically-engineered protein that combines the IgG binding domains of both Protein A and Protein G. The fusion protein is expressed in E. coli. Protein A/G contains four Fc binding domains from Protein A and two from Protein G, resulting in a final mass of 50,460 daltons (40 to 45kDa by SDS-PAGE). Protein A/G is not as pH-dependent as Protein A alone, but otherwise has the additive properties of Protein A and G. Protein A/G binds to all human IgG subclasses, making it the ideal choice for purification of polyclonal or monoclonal IgG antibodies whose subclass identities have not been determined.
Pierce Protein A/G Plus Agarose is prepared using Thermo Scientific AminoLink Coupling Chemistry, which provides several advantages compared to traditional methods of ligand immobilization. AminoLink Immobilization results in conjugation between sugar monomers of the agarose beads and native lysine residues on the Protein A via simple amide bonds. Unlike typical cyanogen bromide (CNBr) immobilization, the AminoLink Method does not introduce any novel chemical groups that could cause undesired nonspecific binding and produces a stable, essentially irreversible bond. The result is a high-binding-capacity resin that retains functional immobilized Protein A/G through multiple rounds of antibody purification.
Properties of crosslinked 6% beaded agarose (CL-6B):
- Support pH Stability: 2 to 14 (short term); 3 to 13 (long term)
- Average Particle Size: 45 to 165 microns
- Exclusion Limit: 10,000 to 4,000,000 daltons
- Maximum Volumetric Flow Rate: approx. 1mL/minute (for 1cm diameter column)
- Maximum Linear Velocity: 30cm per hour
- Maximum Pressure: less than 25psi (1.5 bar), defined as the maximum pressure drop across a column that the resin can withstand (Note: The indicated gauge pressure of a liquid chromatography apparatus may be measuring the total system pressure rather than the pressure drop across the column.)
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