HisTrap excel

• Cat#No#: His-113P
• Product Overview: HisTrap excel columns are prepacked with Ni Sepharose excel affinity resins for capture and purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants by immobilized metal ion affinity chromatography (IMAC).
• Description: HisTrap excel 1 mL and 5 mL are ready-to-use IMAC columns prepacked with Ni Sepharose excel. The design of the columns in combination with the specific properties of the resin enables fast and convenient purifications. The special type of filter in the top and bottom of the columns makes it possible to load large volumes of cell-free, unclarified samples directly on the columns without causing back pressure problems. This time-saving property helps prevent degradation and loss of sensitive target proteins.
• Characteristic: Load eukaryotic cell culture samples containing secreted histidine-tagged proteins directly with retained binding capacity.
  Increase target protein yield and decrease degradation through reduced and simplified sample handling.
  HisTrap excel columns allow direct purification of cell-free, unclarified samples.
• Maximum operating pressure: 5 bar [0.5 MPa] (70 psi)
• Metal ion capacity: 54 to 70 μmol Ni2+ /ml medium
• Matrix: Highly cross-linked spherical agarose, 6%
• Average particle size: 90 μm
• Dynamic binding capacity: At least 10 mg histidine-tagged protein/ml sedimented medium.
• Recommended flow rate: 150 to 600 cm/h
• Recommended column height: 25 mm
• Chemical stability: 0.01 M HCl and 0.01 M NaOH. 10 mM EDTA, 5 mM DTT, 5 mM TCEP, 20 mM β-mercaptoethanol, 1 M NaOH, and 6 M guanidine-HCl. 500 mM imidazole and 100 mM EDTA. 30% 2-propanol.
• Chemical compatibility: Stable in all buffers commonly used in IMAC.
• pH working range: 2 to 12
• CIP stability: 2 to 14
• Storage: 4 to 30°C, 20% ethanol
• Cleaning-in-place: Ionically bound proteins: 1. Wash with several column volumes (CV) of 1.5 M NaCl. 2. Wash with approximately 10 CV distilled water or equilibration buffer.
  Precipitated proteins, hydrophobically bound proteins, and lipoproteins: 1. Wash the column with 1 M NaOH, contact time usually 1 to 2 h (12 to 24 h for endotoxin removal). 2. Wash with approximately 10 CV equilibration buffer.
  Hydrophobically bound proteins, lipoproteins, and lipids: 1. Wash with 5 to 10 CV 30% isopropanol for about 15 to 20 min. 2. Wash with approximately 10 CV distilled water or equilibration buffer. OR 1. Wash with 2 CV 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 h. 2. Remove residual detergent by washing with at least 5 CV 70% ethanol. 3. Wash with approximately 10 CV equilibration buffer.
• Pack size: 5 × 1 mL
• Maximum flow velocity: 600 cm/h
• Wash buffer: 20 mM sodium phosphate, 0.5 M NaCl, 0 to 30 mM imidazole, pH 7.4.
• Dimensions: 7 × 25 mm
• Column volume: 1 ml
• Column i.d.: 7 mm
• Column hardware pressure limit: 5 bar (0.5 MPa)