The Thermo Scientific NAb Protein A/G Spin Kit facilitates rapid, small-scale profiling of antibodies in a variety of sample types. and purification. Each kit includes sufficient NAb Protein A/G spin columns, collection tubes, buffers, and simplified protocols for purification of at least 2 antibody samples.

Pierce Protein A/G Sepharose Features:

•Protein A/G – Immobilized recombinant fusion of the antibody binding domains of Protein A and Protein G Protein enables polyclonal IgG purification of virtually any mammalian species
• Agarose Resin – The support is cross-linked 6% beaded agarose (CL-6B), which is a protein affinity The most commonly used resin for and purification methods
• Inert and Stable – Superior production method immobilizes Protein A/G via uncharged leach-resistant covalent bonds, resulting in lower non-specific binding and Able to be used multiple times without loss of yield
• Standard Capacity – Pierce Protein A/G Agarose contains normal loading amounts of immobilized Protein A/G, providing >7 mg human IgG/ Binding Capacity of mL Resin

Pierce Protein A/G Sepharose contains purified Protein A/G recombinant fusion protein covalently immobilized to high quality cross-linked 6% beaded agarose (CL-6B). This particular resin class provides a versatile combination of chromatographic features for high yields and high-purity purification of whole IgG from mammalian serum samples. The physical and chemical properties of agarose beads lend themselves to many affinity purification systems.

Protein A/G is a genetically engineered protein that combines the IgG binding domains of protein A and protein G. The fusion protein was expressed in E. coli. Protein A/G contains four Fc-binding domains of Protein A and two Fc-binding domains of Protein G, with a final molecular weight of 50,460 daltons (40 to 45 kDa measured by SDS-PAGE). Protein A/G has a lower pH dependence than Protein A alone, but shares the additive properties of Protein A and G.

Protein A/G binds to all human IgG subclasses and is therefore ideal for purification of polyclonal or monoclonal IgG antibodies of unidentified subclasses. In addition, it binds IgA, IgE, IgM and IgD (the latter is weakly binding). Protein A/G also binds to all mouse IgG subclasses but not to mouse IgA, IgM or serum albumin. Therefore, Protein A/G is an excellent choice for purifying and detecting monoclonal antibodies of mouse IgG subclasses, avoiding the interference of IgA, IgM and mouse serum albumin. Individual subclasses of mouse monoclonals are likely to have stronger affinity for chimeric protein A/G than for protein A or protein G.

Pierce Protein A/G Agarose is prepared using Thermo Scientific AminoLink coupling chemistry, which offers several advantages over traditional ligand immobilization methods. After AminoLink immobilization, the sugar monomers of the agarose beads bind to the native lysine residues on protein A through simple amide bonds. Unlike typical cyanogen bromide (CNBr) immobilization methods, the AminoLink method does not introduce any new chemical groups, thereby avoiding possible non-specific binding and forming a stable binding that is essentially irreversible. The resulting high-binding resin retains functional immobilized protein A/G after multiple rounds of antibody purification.

Pierce Protein A/G Agarose binds IgG from nearly all isotypes and mammalian species in serum, ascites fluid, cell culture supernatants, and other antibody samples, enabling efficient antibody purification. Because immobilized Protein A/G combines the immunoglobulin binding domains of Protein A and Protein G, this resin is effective for affinity purification of IgG antibodies from a variety of mammalian host species.

Properties of Cross-Linked 6% Beaded Agarose (CL-6B):
•Supports 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 volume flow rate: approximately 1 mL/min (for 1 cm diameter column)< br>•Maximum linear velocity: 30 cm/h
•Maximum pressure: less than 25psi (1.5 bar); defined as the maximum pressure drop across the column that the resin can withstand (Note: The indicated gauge pressure of the LC unit may Measures total system pressure, not pressure drop across column )

More Product Data
• Antibody purification using antibody binding proteins

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