Benzamidine Sepharose 6B

• Cat#No#: Be-346C
• Product Overview: Benzamidine Sepharose 6B with immobilized p-aminobenzamidine (pABA) is used for removal and/or purification of serine poteases e.g thrombin.
  For convenient, one-step removal and/or purification of trypsin, trypsin-like serine proteases, and zymogens including urokinase and prekallikrein.
  Sepharose 6B, is 6% cross-linked agarose resin. Established resin for purifying serine proteases, e.g. trypsin or thrombin.
  Effective removal of thrombin and factor Xa after tag cleavage of recombinant proteins.
• Description: Benzamidine Sepharose 6B is p-aminobenzamidine covalently attached to Sepharose 6B by the epoxy coupling method. p-Aminobenzamidine (PAB), is a synthetic inhibitor of trypsin-like serine protease. Trypsin and trypsin-like serine proteases bind to Benzamidine Sepharose 6B and can thus be used for purification and/or removal of these substances. Trypsin, bovine thrombin, urokinase, human enterokinase, acrosin, native plasminogen, kallikrein, prekallikrein, collagenase and clostripain are some of the serine proteases that have been purified on Benzamidine Sepharose 6B. For recombinant purification, Benzamidine Sepharose 6B can be used for removal of serine proteases such as thrombin and enterokinase after cleavage of purification tags.
• Applications: Trypsin and trypsin-like serine proteases bind to Benzamidine Sepharose 6B and can thus be used for purification and/or removal of these substances.
• Available capacity: 13 mg trypsin/mL drained media
• Maximum operating pressure: 75 cm/h at 25°C, HR 16/10 column, 5 cm bed height.
• Medium Preparation: Benzamidine Sepharose 6B is supplied pre-swollen in 20% ethanol. Prepare a slurry by decanting the ethanol solution and replacing it with binding buffer in a ratio of 75% settled media to 25% buffer before packing. The binding buffer should not contain agents which significantly increase the viscosity. The column may be equilibrated with viscous buffers at reduced flow rates after packing is completed.
• Matrix: 6% agarose
• Particle Size: 45 µm-165 µm
• Average particle size: ~90 µm
• Ligand: p-aminobenzamidine
• Ligand density: Approx. 7 µmol p-aminobenzamidine/ml drained medium.
• Coupling chemistry: epoxy
• Dynamic binding capacity: 13 mg trypsin/mL drained resin
• Recommended flow rate: 75 cm/h at 25°C, HR 16/10 column, 5 cm bed height.
• Recommended column height: 5 cm
• Chemical stability: Stable to commonly used aqueous buffers.
• Physical stability: Negligible volume variation due to changes in pH or ionic strength.
• pH working range: 3–11
• CIP stability: 2–13
• Storage: 2 to 8°C, 20% ethanol
• Cleaning-in-place: In some applications, substances like denaturated proteins or lipids do not elute in the regeneration procedure. These can be removed by washing the column with a detergent solution, e.g. 0.1% Triton X-100 at 37°C for one minute.
• Pack size: 100 mL