Protein A Sepharose CL-4B antibody purification resin

• Cat#No#: Pr-336C
• Product Overview: Protein A Sepharose CL-4B is a trusted, well-documented affinity resin for immunoprecipitation procedures. It can also be used for antibody purification.
  High binding capacity of 20 mg human IgG/mL.
  High selectivity and low nonspecific adsorption.
  Well suited for immunoprecipitation procedures.
• Description: Protein A Sepharose CL-4B is protein A immobilized by the CNBr method to Sepharose CL-4B.
  Protein A binds to the Fc region of immunoglobulins through interactions with the heavy chain. The binding of protein A has been well documented for IgG from a variety of mammalian species and for some IgM and IgA as well. Protein A Sepharose CL-4B has been used as a powerful tool to isolate and purify classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media. Since only the Fc region is involved in binding, the Fab region is available for binding antigen. Hence, protein A Sepharose CL-4B is extremely useful for isolating of immune complexes.
• Available capacity: ~20 mg human IgG/mL drained medium
• Medium Preparation: Protein A Sepharose CL-4B is supplied preswollen in 20% ethanol. Prepare a slurry by decanting the 20% ethanol solution and replace it with binding buffer in a ratio of 75% settled medium to 25% buffer. The binding buffer should not contain agents which significantly increase the viscosity. The column may be equilibrated with viscous buffers at reduced flow rate after packing is completed.
  For batch procedure remove the ethanol by washing the medium on a medium porosity sintered glass funnel.
• Ligand Coupling Method: Cyanogen bromide activation
• Matrix: Cross-linked agarose, 4%
• Average particle size: ~90 µm
• Ligand: Native Protein A
• Ligand density: 3 mg Protein A/ml medium
• Dynamic binding capacity: ~20 mg human lgG/mL resin
• Recommended flow rate: < 150 cm/h at 25°C, HR 16/10 columns, 5 cm bed height
• Chemical stability: Stable in commonly used aqueous buffers, 6 M guanidine-HCl, 8 M urea.
• Physical stability: Negligible volume variation due to changes in pH or ionic strength.
• pH working range: 3–9
• CIP stability: 2–10
• Storage: 2 to 8°C, 20% Ethanol
• Cleaning-in-place: 1.Remove precipitated or denatured substances by washing the medium with 2 column volumes of 6 M guanidine hydrochloride. Immediately re-equilibrate with at least 5 column volumes of binding buffer.
  2.Remove strongly bound hydrophobic proteins, lipoproteins and lipids by washing the medium with a non-ionic detergent, e.g. 0.1%. Triton X-100, at 37 °C for one minute. Immediately re-equilibrate with at least 5 column volumes of binding buffer.
  3.Alternatively, wash the medium with 70% ethanol and let it stand for 12 hours. Re-equilibrate with at least 5 column volumes of binding buffer.
• Sanitization: Sanitize the packed column with 2% Hibitane/20% ethanol or with 70% ethanol.
• Pack size: 1.5 g
• Maximum flow velocity: 150 cm/h at 25°C, HR 16/10 column, 5 cm bed height