HiBond Ni-NTA Agarose

• Cat#No#: AC-005I
• Product Overview: HiBond Ni-NTA Agarose utilizes 4% agarose as the matrix support, and is functionalized with the chelate ligand Nitilotriacetic Acid (NTA), which is charged with nickel ions (Ni2+) to form a very stable octahedral structure with the nickel ion in the center. The structure of Ni-NTA complex is compatible solutions that contain certain concentration of reducing agents, denaturing agents, detergents and other additives.
  HiBond Ni-NTA Agarose can be used to purify 6xHis-tagged proteins expressed in various hosts, such as E.coli., yeast, insect cells and mammalian cells.
• Characteristic: Suited for routine purification of histidine-tagged proteins
  High yield (typically > 40 mg protein /mL resins)
  Low leakage of nickel ions. Can be re-used for up to 8 preps*
  *Note: No significant decrease in yield after 8 repetitive uses of resin when recommended buffers are used. Refer to Product Details for compatible reagents.
• Applications: Manual batch or gravity flow purification
  Routine His tag protein purification for structural biology and functional assays
  Protein-protein and protein-nucleic acid interaction studies
• Matrix: 4% agarose supplied as 50% slurry
• Average particle size: 45 μm - 165 μm
• Ligand: Nickel
• Dynamic binding capacity: >40 mg 6xHis-tagged protein/mL medium
• Recommended flow rate: Gravity flow, <75 cm/hour
• Maximum Pressure Drop: 0.1 MPa (1 bar)
• Chemical compatibility: (1) Reducing agents
  5 mM DTE
  0.5-1 mM DTT
  20 mM β-mercaptoethanol
  5 mM TCEP
  10 mM reduced glutathione (GSH)
  (2) Denaturing agents
  8 M urea, 6 M Gua-HCl
  (Tested for one week at 40°C)
  (3) Detergents
  2% Triton X-100 (nonionic)
  2% Tween 20 (nonionic)
  2% NP-40 (nonionic)
  2% cholate (anionic)
  1% CHAPS (zwitterionic)
  (4) Other additives
  500 mM imidazole
  20% ethanol
  50% glycerol
  100 mM Na2SO4
  1.5 M NaCl
  1 mM EDTA
  60 mM citrate
  (5) Commonly used buffer
  50 mM sodium phosphate, pH 7.4
  100 mM Tris-HCl, pH 7.4
  100 mM Tris-acetate, pH 7.4
  100 mM HEPES, pH 7.4
  100 mM MOPS, pH 7.4
  100 mM sodium acetate, pH 4
  100 mM MOPS, pH 7.4
  100 mM sodium acetate, pH 4
• Storage: 2 - 8°C, storage buffer: 1X PBS containing 20% ethanol