Ni Sepharose 6 Fast Flow resin

• Cat#No#: Ni-401C
• Product Overview: Ni Sepharose 6 Fast Flow is a high binding-capacity IMAC chromatography resin for purifying his-tagged proteins. Good choice for purification at a variety of scales.
• Description: Purifying histidine-tagged recombinant proteins by immobilized metal affinity chromatography (IMAC) continues to grow in popularity. Nickel (Ni2+) is the most commonly used metal ion in IMAC purifications. Ni Sepharose 6 Fast Flow is a BioProcess medium (resin) that combines the advantages of using Ni2+ for purification of histidine-tagged proteins with the well-established properties of the Sepharose Fast Flow platform.
• Characteristic: Fast, reliable scale-up of histidine-tagged protein purifications.
  High protein binding capacity, and minimal leakage of Ni2+ ions.
  Compatible with a very wide range of reducing agents, detergents, and other additives.
  As a BioProcess medium, Ni Sepharose 6 Fast Flow meets industrial demands with security of supply and comprehensive regulatory support.
  Available as prepacked HisPrep and HisTrap columns for added speed, convenience, and reproducibility.
  Suitable for gravity-flow purification using His GraviTrap columns, and multiwell plate screening using His MultiTrap plates.
• Maximum operating pressure: 0.1 MPa, 1 bar (when packed in XK columns. May vary if used in other columns).
• Sample preparation: The sample should be fully dissolved. To avoid column clogging, we recommend centrifugation and filtration through a 0.45 μm filter to remove cell debris or other particulate material. If the sample is dissolved in a buffer other than 20 mM phosphate buffer with 0.5 M NaCl pH 7.4, adjust its NaCl concentration to 0.5 M and pH to 7–8. Do not use strong bases or acids for pH-adjustments (precipitation risk).
• Metal ion capacity: ~15 µmol Ni2+/ml medium
• Matrix: 6% cross-linked agarose
• Particle Size: 45 µm-165 µm
• Average particle size: ~90 µm
• Dynamic binding capacity: Approx. 40 mg histidine-tagged protein.
• Recommended flow rate: 150 cm/h
• Chemical stability: Stable in commonly used aqueous buffers - 0.1 M HCl, 0.1 M NaOH, 8 M urea
• Chemical compatibility: Stable in all commonly used buffers, reducing agents, denaturants, and detergents.
• pH working range: 3–12
• CIP stability: 2–14
• Temperature stability: 4ºC to 30ºC
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: 1. Wash with several column volumes of 1.5 M NaCl. Then wash with several column volumes of distilled water.
  2. Wash the column with 1 M NaOH, contact time usually 1 to 2 hours (12 hours or more to remove endotoxins). Then wash with approximately 10 column volumes of binding buffer, followed by 10 column volumes of distilled water.
  3. Wash with 5 to 10 column volumes of 30% isopropanol for about 15 to 20 minutes. Then wash with approximately 10 column volumes of distilled water.
  4. 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 5 to 10 column volumes of 70% ethanol1 . Then wash with approximately 10 column volumes of distilled water.
• Pack size: 5 mL
• BioProcess resin: Yes
• Maximum flow velocity: 600 cm/h (20 mL/min) using XK 16/20 column with 5 cm bed height
• Dimensions: 0.7 × 2.5 cm (1 mL column)
• Column volume: 1 mL
• Column hardware pressure limit: 5 bar (0.5 MPa, 73 psi)