Protein G Sepharose 4 Fast Flow resin

• Cat#No#: Pr-337C
• Product Overview: Protein G Sepharose Fast Flow is recombinant protein G coupled to Sepharose 4 Fast Flow resin.
  High binding capacity: approximately 20 mg human IgG per mL resin.
  Broad-spectrum: binds to a broad range of IgG species and subclasses.
  Complements Protein A resins: unlike protein A, protein G binds all human and mouse IgG subclasses, including human IgG3 and mouse IgG1 as well as all rat IgG subclasses.
  No specific albumin binding: recombinant protein G has been genetically engineered to remove the albumin binding region.
  Minimal leaching: optimized coupling method minimizes leaching of recombinant protein G and improves binding capacity for IgG.
  Wide range of formats: including loose resin packs from 5 mL to 5 L, for manual or automated antibody purification and immunoprecipitation procedures.
• Characteristic: Protein G is immobilized by the well-documented CNBr method on Sepharose 4 Fast Flow, a cross-linked 4% agarose derivative with unique chemical and physical stability. The kinetics of the matrix impart excellent chromatographic properties to the affinity adsorbent, which ensures high yields of separated IgG.
• Applications: Isolation and purification of IgG from serum, ascites, and tissue culture media.
  Monoclonal antibody purification.
  Isolation of antibody-ligand immune complexes.
  Immunoprecipitation protocols.
  Protein G Sepharose Fast Flow resin is used in specific separation applications where broad binding specificity is required.
• Maximum operating pressure: < 0.1 MPa in a XK 50/60 column with 5 cm diameter and 25 cm bed height (at 20˚C using buffers with the same viscosity as water).
• Ligand Coupling Method: Cyanogen bromide activation
• Packing Column: To pack the resin, first wash away the ethanol solution with distilled water (pH 7) on a sintered glass filter or similar. While the resin is still on the filter, resuspend it in binding buffer (e.g., 20 mM phosphate buffer [ph 7.0]), and transfer it to the column. Pack the column.
• Matrix: cross-linked agarose, 4%, spherical
• Swelling: Protein G Sepharose 4 Fast Flow is supplied pre-swollen in 20% ethanol.
• Average particle size: ~90 µm
• Ligand: Recombinant protein G lacking albumin-binding region, produced in E.coli.
• Ligand density: 2 mg Protein G/ml medium
• Dynamic binding capacity: ≥ 20 mg human IgG/mL resin
• Recommended flow rate: 150 to 250 cm/h in a XK 50/60 column with 5 cm diameter and 25 cm bed height (at 20˚C using buffers with the same viscosity as water).
• Recommended column height: 25 cm
• Chemical stability: Stable in aqueous buffers commonly used in Protein G chromatography, 6 M guanidine hydrochloride pH 4.7, 20 mM sodium phosphate + 1% SDS, 70% Ethanol, 95% Ethanol, 20 mM Sodium phosphate, 100 mM Glycine- phosphoric acid, 6 M Urea, 20% ethanol + 2% hibitane. digluconate
• Physical stability: Negligible volume variation due to changes in pH or ionic strength.
• pH working range: 3–9
• CIP stability: 2–10
• Temperature stability: 2˚C to 40˚C
• Storage: 2 to 8°C, 20% Ethanol
• Cleaning-in-place: Remove strongly bound hydrophobic proteins, lipoproteins and lipids by washing the column with a nonionic detergent, 0.1%, at 37°C, contact time one minute. Immediately reequilibrate with at least 5 bed volumes of sterile filtered binding buffer. Alternatively, wash the column with 70% ethanol and let stand for 12 hours. Re-equilibrate with at least 5 bed volumes of sterile binding buffer.
• Sanitization: Sanitize the column with 70% ethanol
• Pack size: 5 mL
• Maximum flow velocity: 250 cm/h
• Dimensions: 5 cm