Protein A High-Capacity Agarose and Kits

Thermo Scientific Pierce Protein A Plus Agarose is a versatile, high-performance affinity resin for antibody purification that is available as bottled agarose beads, pre-packed spin columns, 96-well filter plates and complete IgG purification kits.

Pierce Protein A Plus Agarose consists of purified native Protein A that has been covalently immobilized at high density onto high-quality crosslinked 6% beaded agarose (CL-6B). Among the many available forms of immobilized Protein A affinity resin, this premiere variety provides the most versatile combination of chromatographic features for high yield and high purity purification of whole IgG from mammalian serum samples. The agarose beads have physical and chemical properties suitable for many affinity purification systems. Accordingly, Pierce Protein A Plus Agarose is offered in several convenient package formats, including bottled resin slurries, three sizes of spin columns, complete purification kits, two sizes of FPLC-ready chromatography cartridges, and 96-well filter plates.

Highlights:

• 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
• High capacity – this “Plus” variety of Pierce Protein A Agarose has a dense load of immobilized Protein A, providing a binding capacity greater than 34mg human IgG/mL resin (approx. 16 to 17mg mouse IgG/mL resin)
• Multiple formats – choose from bottled resin slurries, centrifuge-ready columns, complete purification kits, two sizes of FPLC-ready chromatography cartridges, and 96-well filter plates

Includes:

• Purification kits contain Protein A Plus Spin Columns, binding buffer, elution buffer and neutralization buffer

Product Details:

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 binds weakly to mouse IgG1 and is not recommended for most applications with that antibody isotype.

Pierce Protein A 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 through multiple rounds of antibody purification.

Using antibody-binding proteins for antibody purification – Choose a column format based on the specific application and scale

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.)

References:

1. Sjoquist, J., et al. (1972) Eur. J. Biochem.29, 572-578.
2. Hjelm, H., et al. (1975) Eur. J. Biochem.57, 395-403.
3. Sjoholm, I., et al. (1975) Eur. J. Biochem.51, 55-6.
4. Wang, B., et al. (1999) Proc. Nat. Acad. Sci. USA.96, 1627-1632.
5. Higashi, I. , et al. (2000) Clin. Chem. 46, 297-299.

Related Resources:

Review of antibody purification methods
Compare Protein A, Protein G and other antibody binding proteins
Protein A, Protein G, Protein L affinity by species and antibody type (Tech Tip #34)

Related Products:

Other Protein A Resins
All Antibody Purification Products
Pierce Protein A/G Magnetic Beads
MagnaBind Protein A Magnetic Beads
Immunoprecipitation Kits

Available Formats:

• Resin Slurries – 1mL, 5mL, 25mL bottles
• Spin Columns – 0.2mL (microcentrifuge), 1mL, 5mL columns
• Purification Kits – complete kits in all three column sizes
• Chromatography Cartridges – compatible with automated liquid chromatography systems and with syringe processing
• 96-well Spin Plates – pre-dispensed 96-well filter plates, compatible with centrifugation and vacuum manifold systems for manual or automated purification. The plates enable fast, consistent well-to-well and plate-to-plate reproducibility for small-scale, high-throughput separations. (see Protein A 96-well Spin Plate data)
• Bulk and Custom – let us know your needs for bulk Pierce Protein A Plus Agarose