NHS-Activated Sepharose 4 Fast Flow

• Cat#No#: NHS-376C
• Product Overview: NHS-activated Sepharose 4 Fast Flow is a pre-activated agarose matrix that increases the choice of coupling chemistries available. Suitable for coupling amino-containing smaller proteins and peptides.
  NHS pre-activated resin for coupling of small amino-containing proteins and peptides in process-scale applications.
  High level of pre-activation gives a high degree of substitution of the selected ligand.
  NHS coupling method results in chemically stable ligand attachment.
  BioProcess resin supported for industrial applications.
• Description: The preparation and use of affinity chromatography resins by coupling group-specific ligands to preactivated resins is a widely used, successful, and well-documented technique. NHS-activated Sepharose 4 Fast Flow is a preactivated agarose matrix that increases the choice of coupling chemistries available. NHS (N-hydroxysuccinimide) coupling forms a chemically stable amide bond with ligands containing primary amino groups. NHS-activated Sepharose 4 Fast Flow provides a spacer arm and is therefore particularly suitable for immobilizing small protein and peptide ligands.
• Characteristic: Particularly suitable for coupling of small amino-containing proteins and peptides in process-scale applications.
  High level of preactivation, which gives a high degree of substitution of the selected ligand.
  NHS coupling method results in chemically stable ligand attachment.
  BioProcess resin supported for industrial applications.
• Maximum operating pressure: 150–250 cm/h at 0.1 MPa (1 bar, 14.5 psi) in an XK 50/60 column, 25 cm bed height (at 25°C using water).
• Matrix: 4% cross-linked agarose
• Particle Size: 45 μm-165 μm
• Average particle size: ~90 µm
• Ligand: N-hydroxysuccinimide (NHS) activated
• Ligand density: ~16-23 µmol NHS/ml drained medium
• Recommended flow rate: 150 cm/h at 100 kPa
• Recommended column height: 25 cm
• Chemical stability: Stable in commonly used aqueous buffers provided that the ligand withstand the pH or the chemical environment.
• pH working range: 2–9
• CIP stability: 2–13
• Storage: 2 to 8°C, 100% Isopropanol
• Cleaning-in-place: Precipitated or denatured substances:Wash with 4 column volumes of 0.1 M NaOH 1 to 2 h contact time. Wash with at least 5 column volumes of sterile filtered binding buffer. Wash with 2 column volumes of 6 M guanidine hydrochloride. Wash substances immediately with at least 5 column volumes of sterile filtered binding buffer.
  Hydrophobically bound substances:Wash the column with 2 column volumes of a non-ionic detergent (conc. 0.1 to 0.5%). Wash immediately with at least 5 column volumes of sterile filtered binding buffer. Wash the column with 3 to 4 column volumes of 70% ethanol. Wash immediately with at least 5 column volumes of sterile filtered binding buffer.
• Sanitization: If ligand stability permits, a recommended sanitization procedure is to equilibrate the packed column with 0.1 M NaOH in 20% ethanol and allow to stand for 1 hour. Wash with at least 5 column volumes of sterile binding buffer. or Equilibrate with a buffer consisting of 2% hibitane digluconate and 20% ethanol. Allow to stand for 6 hours, then wash with at least 5 column volumes of sterile binding buffer. or Equilibrate with 70% ethanol. Allow to stand for 12 hours, then wash with at least 5 column volumes of sterile binding buffer.
• Pack size: 25 mL
• Group To be Coupled: Primary amines