High capacity purification of IgGs and Fc-tagged proteins. Suitable for polyclonal IgG from rabbit, pig, dog and cat serum. >40 mg human IgG/mL resins ... Read more
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.
- 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.
|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|
||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|
Long term: pH 2 - 9;
Short term: pH 2 - 11
||2 - 8°C|
Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.
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