HiBond Ni-NTA Agarose 6FF

• Cat#No#: AC-004I
• Product Overview: HiBond Ni-NTA Agarose 6FF has been optimized for automated purification applications. It has been widely used in scale-up and preparative purification of histidine-tagged proteins that are expressed in various hosts, such as E.coli, yeast, insect cells, and mammalian cells.
  The base matrix of HiBond Ni-NTA Agarose 6FF is highly crosslinked 6% agarose, offering excellent chemical-, and mechanical stability for automated applications. The matrix support is functionalized with the chelate ligand Nitilotriacetic Acid (NTA) in complex with the nickel ions (Ni2+) in a very stable octahedral structure. The structure of Ni-NTA complex is compatible solutions that contain certain concentration of reducing agents, denaturing agents, detergents and other additives.
  The NTA ligand coordinates the Ni2+ with four valencies (tetradentate, coordination number 4), and two valencies are available for interaction with imidazole rings of histidine residues. The number of available valencies seems to provide the best combination of binding effectiveness and chemical stability for binding with the His tag. The Ni-NTA ion-chelate complex on our HiBond Ni-NTA resins has been delicately optimized to ensure the most effective capture for His tag purification of His-tagged proteins.
• Characteristic: Mechanical stable, designed for scaling up His tag purification
  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: Automated or syringe operations; and manual batch or gravity flow
  Routine His tag protein purification for structural biology and functional assays
  Scaling up and preparative applications
• Matrix: 6% highly cross-linked agarose supplied as 50% slurry
• Average particle size: 45 μm - 165 μm
• Ligand: Nickel
• Dynamic binding capacity: > 40 mg 6xHis-tagged protein/mL medium
• Maximum Pressure Drop: 0.3 MPa (3 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
• Storage: 2 - 8°C, storage buffer: 1X PBS containing 20% ethanol