GSTrap 4B Columns

• Cat#No#: GST-097P
• Product Overview: GSTrap 4B columns are prepacked Glutathione Sepharose 4B columns for convenient, high capacity one-step purification of glutathione S-transferase (GST) tagged proteins.
• Description: GST-tagged proteins can be purified directly from pretreated bacterial lysates using GSTrap 4B. GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods. The resin, Glutathione Sepharose 4B, is also available as lab packages and is a good choice for scale-up. The columns can be operated with a syringe, peristaltic pump, or liquid chromatography system.
• Characteristic: High binding capacity.
  Mild elution conditions preserving protein antigenicity and function.
  Easy one-step purifications of Glutathione S-Transferase (GST) tagged proteins resulting in high purity.
  GSTrap 4B are designed to be used with a syringe, pump, or chromatography system, such as ÄKTA design.
• Applications: GSTrap 4B columns are used for the purification of GST-tagged proteins from bacterial lysates and other glutathione S-tranferases or glutathione-dependent proteins. Mild elution conditions preserve protein antigenicity and function.
• Maximum operating pressure: 5 bar [0.5 MPa] (70 psi)
• Sample preparation: The sample should be centrifuged and/or filtered through a 0.45 μm filter immediately before it is applied to the column. If the sample is too viscous, dilute it with binding buffer to prevent clogging the column. Less protein may bind to the medium due to a lower protein concentration in the sample.
• Matrix: 4% agarose
• Average particle size: 90 μm
• Ligand: glutathione and 10-carbon linker arm
• Ligand density: 7 to 15 μmol glutathione/ml medium
• Dynamic binding capacity: Approx. 25 mg recombinant glutathione Stransferase (Mr 26 000)/ml medium (protein dependent).
• Recommended flow rate: < 4 ml/min
• Recommended column height: 25 mm
• Chemical stability: All commonly used aqueous buffers, e.g., 1 M acetate pH 4.0 and 6 M guanidine hydrochloride for 1 h at room temperature.
• CIP stability: 4 to 13
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: Removal of precipitated or denatured substances: Wash with 2 column volumes of 6 M guanidine hydrochloride, immediately followed by 5 column volumes of PBS.
  Removal of hydrophobically bound substances: Wash with 3 to 4 column volumes of 70% ethanol or 2 column volumes of 1% Triton X-100 immediately followed by 5 column volumes of PBS.
• Pack size: 5 × 1 mL
• Dimensions: 7 × 25 mm
• Column volume: 1 ml
• Column i.d.: 7 mm
• Column hardware pressure limit: 5 bar (0.5 MPa)