HiTrap IMAC Sepharose FF

• Cat#No#: Hi-077P
• 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.
  Optimal bead size for scaling up.
  High binding capacity.
• Maximum operating pressure: 5 bar [0.5 MPa] (70 psi)
• Sample preparation: Adjust the sample to the composition and pH of the binding buffer by adding buffer, NaCl, imidazole, and additives (as required) from concentrated stock solutions, by diluting the sample with binding buffer, or by buffer exchange. Do not use strong bases or acids for pHadjustment (precipitation risk). Shortly before applying the sample to the column, centrifuge it and/or filter it through 0.45 or 0.22 μm filters.
• Metal ion capacity: Approx. 15 μmol Ni2+/ml medium.
• Matrix: Highly cross-linked spherical agarose, 6%
• 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: 1 ml/min and 5 ml/min for 1 ml and 5 ml column, respectively.
• Chemical stability: 0.01 M HCl, 0.1 M NaOH. Tested for 1 week at 40°C. 1 M NaOH, 70% acetic acid. Tested for 12 hours. 2% SDS. Tested for 1 hour. 30% 2-propanol. Tested for 30 minutes
• pH working range: 2–14
• CIP stability: 3–12
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: Remove ionically bound proteins by washing with several CV of 1.5 M NaCl. Then wash with at least 3 CV of distilled water.
  Remove precipitated proteins, hydrophobically-bound proteins and lipoproteins by washing with 1 M NaOH, contact time usually 1–2 h (longer time may be required to inactivate endotoxins). Then wash with 3–10 CV of binding buffer, followed by 5–10 CV of distilled water.
  Remove hydrophobically bound proteins, lipoproteins, and lipids by washing the column with 5–10 CV 30% isopropanol for at least 15–20 min. Then wash with approx. 10 CV of distilled water.
• Pack size: 5 × 1 mL
• Maximum flow velocity: 4 ml/min and 20 ml/min for 1 ml and 5 ml column, respectively.
• Column volume: 1 mL
• Column i.d.: 7 mm
• Column hardware pressure limit: 5 bar (0.5 MPa, 70 psi)