Description
GoldBio’s Glutathione Agarose Resin is an affinity chromatography resin that provides a rapid, mild, and highly selective one-step purification method for recombinant proteins containing glutathione S-transferase (GST) tags.
Glutathione has been covalently immobilized onto an agarose matrix, creating a highly selective affinity support that binds GST-tagged recombinant proteins while minimizing nonspecific protein interactions.
Because GST binds reduced glutathione with high affinity, immobilization of glutathione on agarose enables efficient one-step purification of GST fusion proteins from crude lysates.
The resin is supplied as a 75% (v/v) aqueous suspension in 20% ethanol and is compatible with gravity-flow columns, batch purification, and other standard affinity chromatography formats.
GST fusion proteins expressed from pGEX and related expression vectors can be purified to high purity under mild conditions, helping preserve protein structure and biological activity. Bound proteins are readily eluted using buffers containing reduced glutathione, allowing efficient recovery while maintaining protein functionality.
Glutathione Agarose Resin is widely used in recombinant protein purification, protein interaction studies, structural biology, enzyme characterization, and proteomics research. The mild purification conditions and high specificity of the glutathione-GST interaction make this resin an excellent choice for isolating functional recombinant proteins suitable for downstream biochemical and analytical applications.
Glutathione Agarose Resin Working Conditions
Table 1: Glutathione Working Conditions
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Volumetric flow rate (mL / min)
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Column diameter
(mm)
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Bed volume
(mL)
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Packing
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Equilibration/Washing/Elution
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Binding
|
|
6.6
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1.0
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1.4
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1.0
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0.3 - 1.0
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|
16
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10
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7.0
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5.0
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0.5 - 5.0
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|
|
|
|
|
|
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Linear flow rate (cm/h)
|
|
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<250
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<180
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<180
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Quick Details
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Bead Geometry & Size
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Spherical ~50 - 150 µm
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Agarose %
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4 %
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Suspension
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75% (v/v) aqueous suspension in 20% ethanol
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|
Static Binding Capacity
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≥ 8 mg recombinant GST / ml gel
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Storage/Handling
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Store at 4°C. Do not freeze.
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Mechanism
Glutathione Agarose Resin functions through affinity chromatography based on the highly specific interaction between immobilized reduced glutathione and glutathione S-transferase (GST).
Recombinant proteins engineered with an N-terminal or C-terminal GST tag selectively bind to the immobilized glutathione ligand while most endogenous proteins are removed during washing. Elution is accomplished by introducing reduced glutathione into the buffer, which competitively displaces the GST-tagged protein from the resin under mild, non-denaturing conditions, helping preserve protein activity and native structure.
Common Applications
(Click each for more information)
Purification of GST Fusion Proteins
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Purpose: To selectively isolate GST-tagged recombinant proteins from complex biological samples.
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How It Works: Immobilized glutathione specifically binds the GST affinity tag, allowing contaminants to be removed before elution with reduced glutathione.
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Applications: Recombinant protein purification, expression optimization, and protein production.
Smith, D. B., & Johnson, K. S. (1988). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene, 67(1), 31–40.
Protein-Protein Interaction Studies
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Purpose: To isolate GST-tagged bait proteins together with interacting binding partners.
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How It Works: GST fusion proteins are immobilized on glutathione resin, allowing associated proteins to be captured and analyzed.
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Applications: GST pull-down assays, interaction mapping, and signaling pathway research.
Kim, S. Y., & Hakoshima, T. (2019). GST Pull-Down Assay to Measure Complex Formations. Methods in Molecular Biology, 1893, 273–280. https://doi.org/10.1007/978-1-4939-8910-2_20
Structural Biology and Functional Protein Analysis
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Purpose: To obtain highly purified proteins for structural and biochemical characterization.
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How It Works: High-affinity purification minimizes contaminating proteins while maintaining native protein conformation.
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Applications: Protein structural studies, enzyme kinetics, biochemical characterization, and functional assays.
Terpe, K. (2003). Overview of tag protein fusions: From molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523–533.
High-Throughput Recombinant Protein Purification
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Purpose: To purify multiple GST fusion proteins efficiently in parallel.
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How It Works: The resin is compatible with batch purification and column formats that support scalable protein purification.
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Applications: Protein expression screening, clone evaluation, and recombinant protein production.
Arnau, J., Lauritzen, C., Petersen, G. E., & Pedersen, J. (2006). Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. Protein Expression and Purification, 48(1), 1–13.
Purification of Active Enzymes Under Mild Conditions
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Purpose: To isolate recombinant enzymes while preserving biological activity.
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How It Works: Elution with reduced glutathione avoids harsh conditions that can denature sensitive proteins.
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Applications: Enzyme characterization, catalytic studies, and functional protein assays.
Harper, S., & Speicher, D. W. (2011). Purification of proteins fused to glutathione S-transferase. Methods in Molecular Biology, 681, 259–280. https://doi.org/10.1007/978-1-60761-913-0_15
Key Benefits
- One-step affinity purification of GST-tagged recombinant proteins.
- High specificity through the glutathione-GST interaction, reducing nonspecific protein binding.
- Mild elution conditions using reduced glutathione help preserve native protein structure and activity.
- Supplied as a ready-to-use 75% (v/v) aqueous suspension in 20% ethanol.
Storage/Handling
Store at 4°C. Do not freeze.
Supplied as a 75% (v/v) aqueous suspension in 20% ethanol.
For research use only. Not for food, drug, household, or cosmetic use.
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