IMAC Sepharose High Performance resin

• Cat#No#: IM-370C
• Product Overview: IMAC Sepharose High Performance is an uncharged IMAC resin for purifying proteins and peptides with an affinity for metal ions: Supplied uncharged, for customized metal ion charging and optimized selectivity.
  Suitable for optimizing purification of histidine-tagged (his-tagged) proteins when nickel is not the best choice of metal ion.
  High performance purification due to its 34 μm bead size.
  High binding capacity.
• Description: Proteins and peptides that have an affinity for metal ions can be purified using immobilized metal ion affinity chromatography (IMAC), a method that has been growing in popularity and effectiveness.
  IMAC Sepharose HP 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. The immobilized metal ions will interact with certain amino acid residues on protein surfaces (mainly histidine, but often also cysteine and tryptophan), if the amino acid side chains are sufficiently exposed. The bound protein can be eluted either with a competitive agent such as imidazole or by lowering the pH.
• Characteristic: Possible to charge with various metal ions for optimized selectivity.
  High protein binding capacity.
  Compatible with a wide range of additives.
  Available in the convenient and time-saving prepacked HiTrap format.
• Maximum operating pressure: 0.3 MPa, 3 bar
• Sample preparation: To avoid column clogging, centrifuge the sample and filter it through a 0.45-μm filter to remove cell debris and other particulate material. If the sample is dissolved in a buffer other than 20 mM phosphate, 0.5 M NaCl, pH 7.4, adjust the NaCl concentration to 0.5 M and the pH to 7–8. This can be done by adding concentrated stock solutions, diluting with binding buffer, or by buffer exchange. Do not use strong bases or acids to adjust pH (risk for precipitation).
• Metal ion capacity: ~15 µmol Ni2+/mL medium
• Packing Column: IMAC Sepharose High Performance is supplied preswollen in 20% ethanol. Prepare a slurry by decanting the 20% ethanol solution and replacing it with distilled water in a ratio of 75% settled medium to 25% distilled water.
• Matrix: Highly cross-linked agarose, 6%
• Average particle size: ~34 μm
• Dynamic binding capacity: At least 40 mg (histidine)6-tagged protein/mL medium (Ni2+-charged)
• Recommended flow rate: < 150 cm/h
• Chemical stability: Stable in commonly used buffers, At 40°C 0.01 M HCl, 0.1 M NaOH [For One Week] 1M NaOH, 70% acetic acid [For 12 h] 30% 2-propanol [30 min] 2% SDS [1 h]
• pH working range: 3–12
• CIP stability: 2–14
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: Remove ionically bound proteins by washing with at least 0.5 column volumes (CV) of 2 M NaCl. Then wash with at least 3 CV of distilled water.
  Remove precipitated proteins, hydrophobically bound proteins and lipoproteins by washing the column with 1 M NaOH, contact time usually 1–2 hours (longer time may be required to inactivate endotoxins). Then wash with at least 3 (up to 10) CV of binding buffer, followed by at least 3 CV of distilled water.
  Remove hydrophobically bound proteins, lipoproteins and lipids by washing with 5–10 CV of 70% ethanol or 30% isopropanol for at least 15–20 min. Then wash with at least 3 (up to 10) CV of distilled water.
• Pack size: 25 mL
• Maximum flow velocity: 4 and 20 mL/min for 1 and 5 mL column, respectively
• Dimensions: i.d. × h: 0.7 × 2.5 cm (1 mL) 1.6 × 2.5 cm (5 mL)
• Column volume: 1 mL or 5 mL
• Column hardware pressure limit: 5 bar (0.5 MPa, 70 psi)