- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Anti-GST Magnetic Beads, Glutathione Coated, 5 µm, High Purity |
| Catalog No. | MAGBEA-0032 |
| Description | Glutathione-functionalized magnetic agarose beads for one-step affinity purification of glutathione S-transferase (GST)-tagged recombinant proteins from bacterial, yeast, insect, and mammalian expression systems. Glutathione (γ-Glu-Cys-Gly) is covalently immobilized on 5 µm cross-linked magnetic agarose beads via a stable linker. GST-tagged proteins bind specifically to immobilized glutathione with moderate affinity (Kd ~0.1-10 µM for GST from Schistosoma japonicum), enabling efficient capture while allowing competitive elution under mild conditions with reduced glutathione (10-50 mM, pH 8.0). The highly cross-linked agarose matrix provides negligible nonspecific binding, high chemical stability, and compatibility with a wide range of buffer conditions. |
| Intended Use | Single-step affinity purification of GST-tagged recombinant proteins; pull-down assays for GST-fusion protein-protein interaction studies (GST pull-down); enzymatic removal of GST tag using site-specific protease (thrombin, Factor Xa, or PreScission protease) while protein is immobilized on beads. |
| Principle / Technology | Glutathione (GSH) covalently immobilized on magnetic agarose matrix; GST enzyme binds GSH specifically with dissociation constant Kd 0.1-10 µM; binding is reversible; competitive elution with free reduced glutathione (10-50 mM) displaces bound GST-fusion protein; on-bead protease cleavage releases target protein without GST tag. |
| Detection Method | Equilibrate beads in binding buffer; add to clarified lysate; incubate 30-60 min at 2-8 °C or 15-30 min at RT; wash 3-5× with binding buffer; elute with 10-50 mM reduced glutathione pH 8.0; dialyze eluate to remove glutathione or use desalting column. |
| Sample Type | Clarified E. coli lysates (primary application); yeast, insect cell, and mammalian cell lysates; in vitro translation reactions expressing GST fusion proteins. |
| Performance Range / Specifications | Binding capacity: 5-10 mg GST-tagged protein per mL settled beads (typical for GST ~26 kDa fusion); purity >90% in single step from E. coli; GST tag ~26 kDa adds appreciable mass to fusion protein. |
| Sensitivity / LOD | Recovery of GST-fusion proteins from bacterial cultures expressing at 1-5 mg/L; detectable by SDS-PAGE from cultures as small as 10 mL. |
| Specificity | Glutathione specifically binds GST enzyme (Schistosoma japonicum, 26 kDa); minimal binding of endogenous glutathione-binding proteins from E. coli; virtually no binding of non-GST-tagged host proteins at recommended NaCl concentration (150-300 mM). |
| Reaction Conditions / Protocol | Binding: 30-60 min incubation with rotation; wash: PBS or Tris-NaCl with 0.1% detergent; elution: 10-50 mM reduced glutathione, pH 8.0, 10-15 min at RT; complete protocol ~1.5-2 hours. |
| Components / Formulation | Glutathione magnetic agarose bead suspension (50% slurry in 20% ethanol), reduced glutathione (lyophilized, 500 mg), 10× PBS, protocol. |
| Storage Conditions | Store beads at 2-8 °C in 20% ethanol; glutathione powder at 2-8 °C or -20 °C; do not freeze beads. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 1 mL settled beads (2 mL slurry), 5 mL, 25 mL. |
| Product Form | Light brown/beige magnetic bead suspension in 20% ethanol. |
| Quality Control | Each lot tested for GST binding capacity (≥8 mg GST per mL settled beads), purity of purification from E. coli lysate (>90%), and glutathione ligand stability (no leaching detected after 24-hr incubation). |
| Key Features | Glutathione affinity for GST-tagged proteins; magnetic agarose matrix for low nonspecific binding; competitive elution under mild conditions; on-bead protease cleavage for tag removal; reusable 5-10 cycles; chemical stability for cleaning with 6 M guanidine. |
| Purity | Glutathione ligand covalently attached; no detectable leaching; endotoxin <0.1 EU/mL. |
| Concentration | 50% slurry; binding capacity 5-10 mg GST/mL settled beads. |
| Activity / Unit Definition | Glutathione-GST binding: Kd ~0.1-10 µM for S. japonicum GST; competitive elution IC50 ~5-10 mM glutathione. |
| Molecular Weight | Reduced glutathione: 307.32 g/mol; GST: ~26 kDa. |
| Source / Origin | Magnetic agarose beads activated with epoxy or NHS chemistry; glutathione ligand covalently coupled. |
| pH Range / Optimal pH | Binding and wash optimal pH 7.3-7.5; compatible pH range 6.5-8.5. |
| Shipping Conditions | Cold pack (2-8 °C); do not freeze. |
| Expiration Date / Stability | 12 months at 2-8 °C; beads stable in 20% ethanol storage buffer. |
| Regulatory / Compliance | For research use only; not for therapeutic or diagnostic applications. |
| Compatibility | Compatible with PBS, TBS, HEPES, and Tris-based buffers with 150-300 mM NaCl. Nonionic detergents up to 1% compatible. Reducing agents: DTT ≤10 mM compatible. Avoid EDTA >5 mM (may strip glutathione through metal chelation effects). GST binding is NOT compatible with SDS or sarkosyl. Magnetic separation on standard magnetic racks. |
| Recommended Buffer System | PBS or Tris-NaCl buffer, pH 7.4. |
| Application Notes / Precautions | Use fresh or freshly thawed reduced glutathione for elution as glutathione oxidizes rapidly in solution (prepare within 1 hour of use, adjust pH to 7.5-8.0). GST tag can be removed by on-bead cleavage with thrombin (overnight, RT) or PreScission protease (4 hours, 4 °C) — wash beads after binding with protease cleavage buffer, add protease, incubate, collect supernatant containing cleaved target protein. Regenerate beads by washing with 3 bed volumes of 0.1 M Tris-HCl + 0.5 M NaCl pH 8.5, followed by 0.1 M sodium acetate + 0.5 M NaCl pH 4.5, then re-equilibrate. Do not dry beads — keep in suspension at all times. |
| Batch-to-Batch Consistency | Binding capacity within ±20% of reference lot; glutathione ligand density within specification; purity verified from standardized E. coli lysate per lot. |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
