Applications:

OnePass™ GST-Tagged Recombinant Protein Purification column consists of Glutathione-Separopore®4B-CL conveniently prepacked column cartridges for low pressure flow chromatography. This product provides a one step purification method and permits rapid, mild and highly selective purifications of proteins containing glutathione binding sequences. Glutathione-Separopore is prepared by covalently coupling glutathione to epoxy-activated highly crosslinked 4% agarose beads to form a stable thioether linkage. The coupling was optimized to give a high binding capacity and could be greater than 5 mg of Glutathione-S-transferase (GST) per ml of wet gel. Reduced glutathione is covalently linked to Separopore® 4B-CL for use in affinity purification of glutathione-S-transferase (GST) and GST-fused proteins. Bound GST–fusion proteins are easily displaced from the resin by elution with buffers containing reduced glutathione. Mild elution conditions preserve protein antigenicity and function. The OnePass™ prepacked Glutathione- Separopore®spin columns simply purification of GST-tagged proteins.

Technical specifications:

• Ligand: Reduced glutathione (GSH)
• Matrix: Separopore® 4B-CL (highly crosslinked agarose beads, 4%)
• Average Particle size: 52 - 165 µm
• Molecular weight range: 6 x 10⁴ – 2 x 10⁷
• Matrix activation: Epoxy
• Matrix spacer: 12 atoms
• Ligand density: 5 – 10 mg GST / ml drained gel
• Binding capacity: 8 mg recombinant GST-tagged protein / ml drained gel
• pH stability: 3 – 12
• Storage: 2 °C – 8 °C

References:

• pCold-GST vector: a novel cold-shock vector containing GST tag for soluble protein production. Protein Expr Purif. (2008) 62: 120-7.
• Glutathione S-transferase can be used as a C-terminal, enzymatically active dimerization module for a recombinant protease inhibitor, and functionally secreted into the periplasm of Escherichia coli. Protein Sci. (1997) 6: 2180-7.
• Vectors for Cu²⁺-inducible production of glutathione S-transferase-fusion proteins for single-step purification from yeast. Yeast. (1994) 10: 441-9.
• Expression and mutagenesis of recombinant human and murine erythropoietins in Escherichia coli. Biochim Biophys Acta. (1995) 1261: 35-43.
• GST fusion vector with caspase-6 cleavage site for removal of fusion tag during column purification. Biotechniques. (2005) 38: 360-364.
• Bacterial production of recombinant human poly(ADP-ribose) glycohydrolase. Protein Expr Purif. (2011) 75: 230-5.
• Expression of lipase-solubilized bovine liver microsomal cytochrome b5 in Escherichia coli as a glutathione S-transferase fusion protein (GST-cyt b5). Protein Expr Purif. (2006) 45: 352-8.
• Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. (1988) 67: 31-40.
• An automated method for high-throughput protein purification applied to a comparison of His-tag and GST-tag affinity chromatography. BMC Biotechnol. (2003) 3: 12.
• A strategy for high-level expression of soluble and functional human interferon alpha as a GST-fusion protein in E. coli. Protein Eng Des Sel. (2007) 20: 201-9.

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Usage: bioWORLD's products are supplied for LABORATORY RESEARCH USE ONLY. The product may not be used as a drug, agricultural or pesticidal product , food additive or as a household chemical.