PROTEINDEX™ rProtein A Agarose 4 Fast Flow

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% highly cross-linked 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 Mechanically stable matrix support, can be used in spin (at up to 1000 g) and automated applications Minimal ligand leakage, can be reused Versatile, ready-to-use with convenient formats   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 with Ig binding domains Binding capacity >40 mg human IgG/mL medium Matrix Cross-linked 4% agarose supplied as a 50% slurry Particle size 45 μm - 165 μm Maximum pressure 0.3 MPa (3 bar) Chemical stability All commonly used aqueous buffers, including 6 M guanidine-HCl and 8 M urea. Recommended flow rate Gravity flow and medium pressure, 30 - 300 cm/hour pH stability Long term: pH 2 - 9; Short term: pH 2 - 11 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