Activated Thiol Sepharose 4B

• Cat#No#: Ac-342C
• Product Overview: Reversible coupling of proteins and large biomolecules with thiol groups to Sepharose 4B via a glutathione spacer arm.
  The ligand is a mixed disulphide formed between 2,2'-dipyridyl disulphide and glutathione coupled to CNBractivated Sepharose 4B.
  Well suited for covalent chromatography of large molecules such as enzymes and nucleic acids.
  Gel also reacts with heavy metal ions and with alkyl and aryl halides.
  Addition reactions occur with compounds containing C=O, C=C, and N=N bonds.
  Separate thiol-containing proteins from non-thiol-containing proteins.
• Matrix: Agarose, 4%
• Particle Size: 45 μm-165 μm
• Average particle size: ~90 µm
• Ligand density: 1 µmol activated thiol/ml medium
• Recommended flow rate: < 75 cm/h at 25 °C, HR 16/10 column, 5 cm bed height
• Chemical stability: Stable to commonly used aqueous buffers and additives like detergents.
• pH working range: 2–8
• CIP stability: 2–8
• Storage: 2 to 8°C
• Cleaning-in-place: Wash with 5 CV of high pH (100 mM Tris-HCl, 500 mM NaCl, pH 8.5) followed by low pH (100 mM sodium acetate, 500 mM NaCl, pH 4.5). Repeat four to five times. Re-equilibrate immediately with binding buffer.
  Remove precipitated proteins with 4 CV of 100 mM NaOH at a low flow rate, followed by washing with 3 to 4 CV of 70% ethanol or 2 M potassium thiocyanate. Alternatively, wash with 2 CV of 6 M guanidine hydrochloride. Re-equilibrate immediately with binding buffer. Remove strongly hydrophobic proteins, lipoproteins and lipids by washing with 3 to 4 CV of up to 70% ethanol or 30% isopropanol. Alternatively, wash with 2 CV of detergent in a basic or acidic solution, e.g., 0.1% nonionic detergent in 1 M acetic acid at a low flow rate, followed by 5 CV of 70% ethanol to remove residual detergent. Re-equilibrate immediately in binding buffer.
• Pack size: 15 g