Ni Sepharose excel resin

• Cat#No#: Ni-403C
• Product Overview: Ni Sepharose excel is a his-tagged protein purification resin for minimized Ni-leakage and maximized protein recovery when samples contain stripping agents.
• Description: Ni Sepharose excel is immobilized metal ion affinity chromatography (IMAC) media (resins) designed for capture and purification of histidine-tagged proteins from various sample types. Nickel ions are very strongly bound to both media, making them especially suitable for purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants. The media significantly simplify and speed up the workflow since they allow direct loading of large sample volumes without removing agents that normally would cause metal ion stripping. The strong nickel ion binding also provides very high resistance to EDTA and reducing agents like DTT.
• 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.
  Resistant to EDTA (100 mM) and reducing agents such as DTT (5 mM).
  Choose between several different formats for screening and preparative purification of histidine-tagged proteins.
• Metal ion capacity: 54 to 70 µmol Ni2+/ml medium
• Packing Column: 1. Resuspend the medium and pour the slurry into the column in a single continuous motion.
  2. Allow the medium slurry to sediment for at least 3 hours.
  3. If using a packing reservoir, disconnect the reservoir and fill the remainder of the column with 20% ethanol.
  4. Mount the adapter and connect the column to a pump. Avoid trapping air bubbles under the adapter or in the inlet tubing.
  5. Immediately open the bottom outlet of the column and set the pump to run at the desired flow rate.
  6. Maintain packing flow rate for at least 3 column volumes (CV) after a constant bed height is reached.
  7. Stop the pump and close the column outlet.
  8. With the adapter inlet disconnected, quickly push the adapter down into the column until it reaches the bed surface and then a further 3 to 4 mm into the medium bed. Lock the adapter at this level.
  9. Connect the column to a pump or a chromatography system and start equilibration. Re-adjust the adapter if necessary.
• Matrix: Highly cross-linked agarose, 6%
• Average particle size: ~90 µm
• Dynamic binding capacity: At least 10 mg histidine-tagged protein
• Recommended flow rate: 150 cm/h - 600 cm/h
• Chemical stability: 0.01 M HCl and 0.01 M NaOH (one week); 10 mM EDTA, 5 mM DTT, 1 M NaOH, 5 mM TCEP, 20 mM β-Mercaptoethanol, and 6 M guanidine-HCl (24 hours); 500 mM imidazole and 100 mM EDTA (2 hours); 30% 2-propanol (20 minutes).
• Chemical compatibility: Stable in all buffers commonly used in IMAC
• pH working range: 2–12
• CIP stability: 2–14
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: 1. Wash with several column volumes (CV) of 1.5 M NaCl.
  2. Wash with approximately 10 CV distilled water or equilibration buffer.
• Pack size: 25 mL
• Maximum flow velocity: 600 cm/h