PROTEINDEX™ rProtein A Agarose, Settled Resin

PROTEINDEX™ rProtein A Agarose 4 Fast Flow is an affinity chromatography medium designed for easy, one-step purification of immunoglobulins, notably IgG isotypes, as well as Fc region-containing fusion proteins. Protein A, a bacterial cell wall protein isolated from Staphylococcus aureus, binds to the Fc region of most immunoglobulins through interactions with the heavy chain. Native Protein A contains five high affinity IgG binding domains and many other domains with unknown functions. The recombinant Protein A is a genetically engineered protein that contains only the five high affinity IgG binding domains. The non-essential domains were removed to minimize nonspecific binding. The recombinant Protein A is produced in E. coli, and it essentially binds to immunoglobulins with the same degree of affinity as the native Protein A. rProtein A is capable of binding with high affinity to the Fc region from IgG of several species including human and rabbit. Enabled by proprietary ProteIndex™ technology, immobilization of recombinant Protein A on 4% agarose resins was very stable and was optimized for high yield in laboratory preparations and scale-up applications.   Product Highlights High capacity, > 40 mg human IgG / mL of settled resins High selectivity for high purity Minimal ligand leakage, can be reused High purity and high capacity of rProtein A 4FF resin. 2 mL mouse ascites fluid was loaded on the column containing 0.5 mL settled rProtein A 4 FF resin. Bound immunoglobulin species were eluted with 100 mM Glycine, pH 3.0. Fractions were analyzed by SDS-PAGE. Lanes: (1) Raw ascites fluid; (2) Flow-through; (3) Elution. Note that, heavy and light chains of IgG were mostly absent from the flow-through, indicating that most immunoglobulins were bound to the column.   Product Details Ligand Recombinant protein A produced in E.coli Binding capacity >40 mg human IgG/mL medium Matrix 4% agarose supplied as 50% slurry Particle size 45 μm - 165 μm Maximum pressure 0.1 MPa (1 bar) Chemical stability All commonly used aqueous buffers, including 6 M guanidine-HCl and 8 M urea. Recommended flow rate Gravity flow, 100 - 300 cm/hour pH stability Long term: pH3 - 9; Short term: pH2 - 10 Storage temperature 2 - 8°C Storage buffer 20% ethanol   Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.          Documents Technical Note 🗎    Comparison of Binding Characteristics of Immunoglobulin-Binding Proteins