CNBr-Activated Sepharose 4B

• Cat#No#: CN-349C
• Product Overview: CNBr-activated Sepharose 4B is a pre-activated resin used for coupling antibodies or other large proteins containing -NH₂ groups to the Sepharose media, by the cyanogen bromide method, without an intermediate spacer arm.
  For immobilizing antibodies or other large proteins containing -NH₂ groups by coupling them to the matrix.
  Over 25 years proven track record for coupling ligands containing primary amino groups.
  Multipoint attachment resulting in a chemically stable product.
  The coupling reaction by the cyanogen bromide method is spontaneous, rapid and easy to carry out.
• Description: CNBr-activated Sepharose 4B is a pre-activated resin used for coupling antibodies or other large proteins containing -NH₂ groups to the Sepharose media, by the cyanogen bromide method, without an intermediate spacer arm. For immobilizing antibodies or other large proteins containing -NH₂ groups by coupling them to the matrix Over 25 years proven track record for coupling ligands containing primary amino groups Multipoint attachment resulting in a chemically stable product The coupling reaction by the cyanogen bromide method is spontaneous, rapid and easy to carry out
• Maximum operating pressure: base matrix 70-140 cm/h, pressure drop cm H₂O/bed height=15 cm, bed height 10 cm, 5 cm i.d.
• Medium Preparation: CNBr-activated Sepharose 4B is supplied lyophilized in the presence of additives. These additives must be washed away at low pH (pH 3) before coupling the desired ligand. The use of low pH (pH 3) preserves the activity of the reactive groups, which otherwise hydrolyze at high pH.
  Weigh out the required amount of powder (1 g lyophilized powder gives about 3.5 mL final volume of medium) and suspend it in 1 mM HCl. The medium swells immediately and should now be washed for 15 minutes with 1 mM HCl on a sintered glass filter (porosity G3). Use approximately 200 mL 1 mM HCl per gram freeze-dried powder, added in several aliquots.
• Matrix: 4% agarose
• Particle Size: 45 μm-165 μm
• Average particle size: ~90 µm
• Ligand density: For an efficient adsorbent, 1 to 10 μmoles ligand per mL medium is recommended. For protein ligands, 5 to 10 mg protein per mL medium is recommended:
  Sample pH should be the same as that of the binding buffer. Filter the sample through a 0.22 μm or 0.45 μm filter to prolong the working life of the medium.
  After the sample has been loaded, wash the medium with binding buffer until the base line is stable.
• Recommended flow rate: base matrix 70-140 cm/h, pressure drop cm H₂O/bed height=15 cm, bed height 10 cm, 5 cm i.d.
• Recommended column height: 10 cm
• Chemical stability: Stable to commonly used aqueous solutions. Can be used with non-ionic detergents, denaturing solvents, e.g. 8 M urea and 6 M guanidine hydrochloride. Stable in organic solvents, such as 50% dimethylformamide and 50% dioxane.
• Temperature stability: Coupling is completed within 2 hours at room temperature, 20°C to 25°C. If cold room temperatures are necessary, coupling can be carried out overnight.
• Autoclavable: Not recommended
• Storage: 2 to 8°C
• Pack size: 15 g
• Column i.d.: 5 cm