HisTrap FF

• Cat#No#: His-110P
• Product Overview: HisTrap FF is a ready-to-use column, prepacked with precharged Ni Sepharose 6 Fast Flow for preparative purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).
• Description: Ni Sepharose 6 Fast Flow consists of 90 µm beads of highly cross-linked agarose, to which a chelating ligand has been immobilized. This chelating ligand is charged with Ni2+ ions, the first-choice metal ion for purifying most histidine-tagged proteins. The negligible leakage of Ni2+ ions from the matrix ensures reliable capture of histidine-tagged proteins in repeated IMAC purifications.
• Characteristic: High binding capacity, approx. 40 mg/mL resin.
  Negligible leakage of Ni2+.
  Prepacked columns offer reliable and convenient time-saving purification of histidinetagged recombinant proteins.
  Compatible with a wide range of reducing agents, detergents, denaturants, and other additives.
• Maximum operating pressure: 5 bar [0.5 MPa] (70 psi)
• Metal ion capacity: ~ 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.
• Recommended flow rate: 1 ml/min and 5 ml/min for 1 ml and 5 ml column, respectively.
• Recommended column height: 25 mm
• Chemical stability: 0.01 M HCl, 0.1 M NaOH; Tested for one week at 40°C. 1 M NaOH, 70% acetic acid; Tested for 12 h. 2% SDS; Tested for 1 h. 30% 2-propanol; Tested for 30 min.
• Chemical compatibility: Stable in all commonly used buffers, reducing agents, denaturants, and detergents.
• pH working range: 2 to 14
• CIP stability: 3 to 12
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: Ionically bound proteins: Wash with several column volumes of 1.5 M NaCl; then wash with approx. 10 column volumes of distilled water.
  Precipitated proteins, hydrophobically bound proteins, and lipoproteins: Wash the column with 1 M NaOH, contact time usually 1 to 2 hours (12 hours or more for endotoxin removal). Then wash with approx. 10 column volumes of binding buffer, followed by 5 to 10 column volumes of distilled water.
  Hydrophobically bound proteins, lipoproteins, and lipids: Wash with 5 to 10 column volumes of 30% iso-propanol for about 15 to 20 minutes. Then wash with approx. 10 column volumes of distilled water. Alternatively, wash with 2 column volumes of detergent in a basic or acidic solution. Use, for example, 0.1 to 0.5% nonionic detergent in 0.1 M acetic acid, contact time 1 to 2 hours. After treatment, always remove residual detergent by washing with at least 5 column volumes of 70% ethanol. Then wash with approx. 10 column volumes of distilled water.
• Pack size: 5 × 5 mL
• Maximum flow velocity: 4 ml/min and 20 ml/min for 1 ml and 5 ml column, respectively.
• Dimensions: 7 × 25 mm
• Column volume: 1 ml
• Column i.d.: 7 mm
• Column hardware pressure limit: 5 bar (0.5 MPa)