Thermo Scientific Pierce Protein A Agarose is a standard-capacity Protein A beaded agarose resin for use in a variety of antibody affinity purification methods.
Pierce Protein A Agarose consists of purified native Protein A that has been covalently immobilized onto high-quality crosslinked 6% beaded agarose. The agarose beads have physical and chemical properties that enable them to be used in a variety of batch- or column-type affinity purification procedure. Several package formats are offered: 5mL and 25mL bottles of resin slurry, 1mL pre-packed columns and a complete IgG purification kit.
- Native Protein A – immobilized Protein A is ideal for polyclonal IgG purification from human, rabbit, pig, dog or cat serum
- 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 by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
- Standard capacity – Pierce Protein A Agarose has a normal load of immobilized Protein A, providing a binding capacity of 12 to 19mg human IgG/mL resin (approx. 6 to 8mg mouse IgG/mL resin)
- Kit contains Protein A Column (1mL each), Protein A IgG Binding Buffer, IgG Elution Buffer and desalting columns
Protein A is a cell wall component produced by several strains of Staphylococcus aureus. The 46.7kDa-protein consists of a single polypeptide chain that is essentially devoid of carbohydrate. Native Protein A has contains four high-affinity (Ka = 10^8/mol) binding sites that are capable of interacting with the Fc region of IgG-class antibodies from selected mammalian species. IgG-binding function is optimal at pH 8.2, but efficient binding also occurs in neutral and physiological buffers (pH 7.0 to 7.6).
Protein A is a bacteria-derived protein that binds with high affinity and specificity to the Fc portion of antibodies, especially those of the IgG class. Compared to its alternative (Protein G), Protein A provides the highest binding capacity for subclasses of IgG from rabbits, pigs, dogs and cats. Protein A also has higher binding capacity for human IgG, except for IgG3, which it binds weakly. Protein A also binds weakly to mouse IgG1 and is not recommended for most applications with that antibody isotype.
Pierce Protein A 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 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.)
Review of antibody purification methods
All Protein A Resins (including Protein A Agarose and IgG Purification Kits)
All Antibody Purification Products
- Resin Slurries – crosslinked 6% beaded agarose; 5mL and 25mL bottles
- Spin Columns – 1mL columns
- Purification Kit – complete kit containing five 1mL columns
- Bulk and Custom – let us know your needs for bulk Pierce Protein A Agarose