Strep-Tactin XT Sepharose

• Cat#No#: St-405C
• Product Overview: A chromatography resin for purification of recombinant proteins tagged with Strep-tag II and Twin-Strep-tag. These proteins bind very specifically to the immobilized Strep-Tactin XT ligand giving highly pure target protein.
• Description: Strep-Tactin XT Sepharose contains the latest Strep-Tactin ligand, XT, which provides higher affinity, resulting in higher yields and lower loss of valuable target protein than with the predecessor product, StrepTactin Sepharose High Performance (HP). Strep-Tactin XT Sepharose also enables purification under denaturing conditions.
• Characteristic: Highly pure Strep-tag II and Twin-Strep-tag recombinant proteins.
  Fast purifications with simple elution.
  Physiological conditions and mild elution preserve target protein activity.
  Fast and easy regeneration.
  Compatible with a wide range of detergents, denaturants, and other additives.
  Available in 10 and 50 mL lab packs of bulk resin and prepacked in 1 and 5 mL StrepTrap XT chromatography columns for convenience.
• Applications: Purification of recombinant proteins tagged with Strep-tag II and Twin-Strep-tag.
• Sample preparation: Adjust the sample to the composition of the binding buffer. For example, dilute the sample with binding buffer or exchange buffer using a column for desalting. To avoid clogging the column, clarify the sample by centrifugation and filtration through a 0.45 μm filter immediately before application.
• Packing Column: 1. Assemble the column and packing reservoir if necessary.
  2. Remove air from the end-piece 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 resin and pour the slurry into the column in a single continuous motion. Pouring the slurry down a glass rod held against the column wall will minimize the introduction of air bubbles.
  4. Immediately fill the remainder of the column, or packing reservoir, with water. Mount the adapter or lid 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. It is recommended to pack Sepharose HP resin in XK or Tricorn columns in a two-step procedure.
  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 the pump and start the flow. Re-adjust the adapter if necessary.
• Matrix: Rigid, cross-linked agarose, spherical
• Average particle size: 34 µm
• Ligand: Strep-Tactin XT
• Ligand density: ~ 5 mg/mL resin
• Dynamic binding capacity: ~ 10 mg protein with Strep-tag II or Twin-Strep-tag/mL of resin.
• Recommended flow rate: 150 cm/h
• Chemical stability: Stable in all commonly used aqueous buffers
• pH working range: 6 to 10
• CIP stability: 6 to 10
• Storage: 2°C to 8°C in 20% ethanol
• Pack size: 10 mL
• Maximum flow velocity: 300 cm/h
• Maximum operating backpressure: 0.3 MPa (3 bar, 43.5 psi)
• 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: 0.5 MPa (5 bar, 70 psi)