GSTrap FF Columns

• Cat#No#: GST-098P
• Product Overview: GSTrap FF columns are prepacked Glutathione Sepharose Fast Flow columns for fast, convenient, one-step purification of glutathione S-transferase (GST) tagged proteins.
• Description: GST-tagged proteins can be purified directly from pretreated bacterial lysates using GSTrap FF.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 4 Fast Flow, is also available as lab packages and is an excellent choice for scale-up.The columns can be operated with a syringe, peristaltic pump, or liquid chromatography system.
• Characteristic: Fast and simple one-step purification of GST-tagged proteins.
  Mild elution conditions preserving protein antigenicity and function.
  Easy scale-up by connecting the columns in series.
  Sepharose 4 Fast Flow provides good flow properties.
• Applications: For the purification of GST-tagged proteins from bacterial lysates and other glutathione S-tranferases or glutathione-dependent proteins and for preparation of cytoplasmic extract.
• Maximum operating pressure: 5 bar [0.5 MPa] (70 psi)
• Sample preparation: The sample should be centrifuged and/or filtered through a 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.
• Matrix: Highly cross-linked 4% agarose
• Average particle size: 90 μm
• Ligand: Glutathione and 10-carbon linker arm
• Ligand density: 120 to 320 μmol glutathione/ml medium
• Dynamic binding capacity: 11 mg GST-tagged protein/ml medium Mr 43 000 (GSTrap FF 1 ml at 1 ml/min).
• 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 hour at room temperature.
• CIP stability: 3 to 12
• 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
• Maximum flow velocity: 4 ml/min and 15 ml/min for 1 ml and 5 ml columns respectively.
• Dimensions: 7 × 25 mm
• Column volume: 1 ml
• Column i.d.: 7 mm
• Column hardware pressure limit: 5 bar (0.5 MPa)