HiPrep IMAC FF Columns

• Cat#No#: Hi-063P
• Product Overview: HiTrap IMAC FF is prepacked with IMAC Sepharose 6 Fast Flow. The resin is charged with the metalion of your choice for Immobilized Metal ion Affinity Chromatography (IMAC) and subsequent purification of polyhistidine tagged proteins.
• Description: IMAC Sepharose FF is supplied free of metal ions. It is charged by the user with the transition metal ion of choice (e.g. Cu2+, Zn2+, Ni2+, or Co2+); these metal ions will bind to the covalently immobilized chelating ligand on the Sepharose.
• Characteristic: For optimizing purification of histidine-tagged proteins when Ni2+ is not the best choice of metal ion.
  Conveniently charge with your metal of choice.
  Prepacked 20 mL HiPrep columns for easy scale-up.
  High binding capacity.
• Maximum operating pressure: 1.5 bar [0.15 MPa] (22 psi)
• Sample preparation: Centrifuge at 10 000 × g (or higher) for 10 min and/or filter the sample through 0.45-μm filter. If possible, dilute the sample in binding buffer. The sample should contain imidazole at the same concentration as in the binding buffer.
• Metal ion capacity: Approx. 15 μmol Ni2+ /ml medium
• Matrix: Highly cross-linked 6% agarose
• Average particle size: 90 μm
• Dynamic binding capacity: Approx. 40 mg (histidine)6- tagged protein/ml medium (Ni2+ - charged). Untagged protein: Approx. 25 mg/ml medium (Cu2+ charged), or approx. 15 mg/ml medium (Zn2+ or Ni2+ charged).
• Recommended flow rate: < 300 cm/h
• Chemical stability: 1 M NaOH, 70% acetic acid. Tested for 12 h. 2% SDS. Tested for 1 h. 30% 2-propanol. Tested for 30 min.
• pH working range: 2–14
• CIP stability: 3–12
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: Removal of ionically bound substances: Wash with approximately 20 ml 1.5 M NaCl. Then wash the column with approximately 200 ml distilled water.
  Removal of precipitated and/or hydrophobically-bound substances and lipoproteins: Wash with 1 M NaOH, contact time usually 1–2 h (longer time may be required to inactivate endotoxins); then wash with approximately 200 ml binding buffer, followed by 100–200 ml distilled water.
  Removal of hydrophobically-bound proteins, lipoproteins, and lipids: Wash with 100–200 ml 30% isopropanol for at least 15–20 min; then wash with approximately 200 ml distilled water. Alternatively, wash with 40 ml detergent in a basic or acidic solution. Use, for example, 0.1–0.5% nonionic detergent in 0.1 M acetic acid, contact time 1–2 h. After treatment, always remove residual detergent by washing with at least 100 ml 70% ethanol. Then wash with approximately 200 ml distilled water.
• Pack size: 20 mL
• Column volume: 20 ml
• Column i.d.: 16 mm
• Column hardware pressure limit: 0.5 MPa, 5 bar, 73 psi