StrepTactin Sepharose High Performance resin

• Cat#No#: St-406C
• Product Overview: StrepTactin Sepharose High Performance resin is designed for Strep-tag II recombinant protein purification.
  Yields high-purity tagged recombinant proteins eluted in concentrated, small volumes.
  Physiological conditions and mild elution preserve target protein activity.
  Fast and easy regeneration with 0.5 M NaOH.
  Compatible with a wide range of reducing agents, detergents, denaturants, and other additives.
  Stable in all commonly used buffers.
• Description: StrepTactin Sepharose High Performance is a chromatography medium for purifying Strep(II)-tagged recombinant proteins. The medium is available in 10 ml and 50 ml lab packs and prepacked in 1 ml and 5 ml StrepTrap HP columns.
  The Strep(II) tag is a small tag consisting of only eight amino acids (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) and a relative molecular mass (Mr ) of only 1000. The small size of the tag is very beneficial, since in most cases it does not interfere with structural and functional studies and, therefore, does not have to be removed from the target protein. The Strep(II) tag binds very specifically to the immobilized StrepTactin ligand giving pure target protein. Affinity purification using StrepTactin Sepharose High Performance takes place under physiological conditions, and mild elution with desthiobiotin preserves the activity of the target protein.
• Characteristic: Highly pure Strep(II)-tagged recombinant proteins eluted in concentrated form and small volumes.
  Physiological conditions and mild elution preserve target protein activity.
  Fast and easy regeneration with 0.5 M NaOH.
  Compatible with a wide range of reducing agents, detergents, denaturants and other additives.
  Prepacked StrepTrap HP 1 ml and 5 ml columns offer convenience, save time, and ensure reproducible results.
  Easy scale-up.
• Sample preparation: Adjust the sample to the composition of the binding buffer. To avoid clogging the column when loading large sample volumes, filter the sample through a 0.45 μm filter or centrifuge it immediately before application.
• Packing Column: 1. Assemble the column (and packing reservoir if necessary).
  2. Remove air from the end-piece and adapter by flushing with water. Make sure no air has been trapped under the column bed support. Close the column outlet leaving the bed support covered with water.
  3. Resuspend the medium and pour the slurry into the column in a single continuous motion.
  4. If using a packing reservoir, immediately fill the remainder of the column and reservoir with water. Mount the adapter or lid of the packing reservoir and connect the column to a pump. Avoid trapping air bubbles under the adapter or in the inlet tubing.
  5. 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 bed volumes after a constant bed height is reached. Mark the bed height on the column.
  7. Stop the pump and close the column outlet.
  8. If using a packing reservoir, disconnect the reservoir and fit the adapter to the column.
  9. With the adapter inlet disconnected, push the adapter down into the column until it reaches the mark. Allow the packing solution to flush the adapter inlet. Lock the adapter in position.
  10. 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: ~34 µm
• Ligand: StrepTactin
• Ligand density: approx. 5 mg StrepTactin/ml medium.
• Dynamic binding capacity: Approx. 6 mg Strep II-tagged protein/mL resin
• Recommended flow rate: < 300 cm/h
• Chemical stability: Stable in commonly used buffers, 0.5 M NaOH (regeneration and cleaning), reducing agents, denaturants, and detergents.
• pH working range: > 7
• Storage: 2 to 8°C, 20% Ethanol
• Cleaning-in-place: 1. Regenerate and clean the column with 3 CV distilled water followed by 3 CV 0.5 M NaOH and 3 CV distilled water.
  2. Re-equilibrate the column with 5 CV of binding buffer before starting the next purification.
• Pack size: 10 mL
• Maximum flow velocity: < 300 cm/h
• Dimensions: 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)