GST-tagged Fusion Protein Purification

Reduced glutathione (GSH), when immobilized as a ligand to agarose or other chromatography supports, enables high-yield and high-quality purification of recombinant proteins expressed as fusions with glutathione S-transferase (GST). GST-fusion proteins are purified with high yield because of the 12-atom GSH linker, which minimizes steric hindrance. We offer glutathione magnetic beads and agarose resins in multiple formats to accommodate a variety of needs, from high- throughput screening, batch, pilot, and process purification. Superflow agarose resins have undergone extensive chemical characterization.

  • More formats—magnetic beads, magnetic agarose, loose resin, spin columns and kits, FPLC cartridges, and 96-well filter plates enable fusion protein purification from microgram to kilogram scales.
  • High performance—resins are designed to maximize protein yield and reduce background.
  • Economical—pricing is similar to or better than other suppliers.

Choose a product for GST-tagged fusion protein purification

Product type Pierce Glutathione Magnetic Agarose Beads Pierce Glutathione Agarose Resin Pierce Glutathione Superflow Resin
Bead or resin size 10–40 μm 45–165 μm 60–160 μm
Static binding capacity ≥12 mg/mL ~40 mg/mL ~30 mg/mL
Dynamic binding capacity* N/A ~10.5 mg/mL ~10 mg/mL
Maximum linear flow rate* N/A 800 cm/hr 1,200 cm/hr
Support Magnetite-embedded 6% beaded agarose 6% beaded agarose Highly crosslinked 6% beaded agarose
No. of reuses 0 5 25
Formats available loose beads loose beads, pre-packed spin columns and kits, chromatography cartridges loose beads
Recommended application scale high-throughput batch batch, pilot batch, pilot, process
Order product(s) 78601 16100 (loose resin)
16109 (1 mL cartridge)
16110 (5 mL cartridge)
16111 (spin plate)

Additional Pierce Glutathione Agarose products

Resin Format Resin per column Column size Collection tube size Cat. No.
Glutathione Spin Column 0.2 mL 0.8 mL Microcentrifuge 16103
Glutathione Spin Column 1 mL 2 mL 15 mL conical 16104
Glutathione Spin Column 3 mL 10 mL 50 mL conical 16105
Glutathione Spin Column Kit 0.2 mL 0.8 mL Microcentrifuge 16106
Glutathione Spin Column Kit 1 mL 2 mL 15 mL conical 16107
Glutathione Spin Column Kit 3 mL 10 mL 50 mL conical 16108

Featured product data

Comparison of protein yield and purity between Pierce Glutathione Magnetic Agarose and products from other suppliers. Samples (0.25 mL) of three proteins (GST-green Renilla luciferase, GST-RalGDS, and GST-Protein A/G) were diluted with 0.25 mL binding buffer and purified manually with 25 µL settled beads. Respective suppliers’ protocols were followed for their buffer compositions and volumes. Repeating with fresh lysate, yields were lower than previous purifications (freezer malfunctioned, lysates may have thawed). Pierce Magnetic Agarose had the highest yield and similar purity compared to beads from the other suppliers.

Large-scale FPLC purification of GST produces >98% purity of target protein. Biomass (170 g) containing overexpressed GST was lysed with 1.7 L of lysis buffer. Then, 0.75 L of lysate was loaded onto equilibrated 200 mL columns (50 mm x 100 mm) of Thermo Scientific Pierce Glutathione Superflow Agarose (left) or Glutathione Sepharose™ 4 Fast Flow (GE Healthcare, Cat. 17-5132-03, right) at a linear flow rate of 30 cm/hour. Columns were washed with binding buffer until the UV280 reached baseline; then bound protein was eluted with elution buffer, and fractions containing purified GST were pooled. Aliquots of load, flow-through, wash, and eluate fractions were separated by SDS-PAGE, stained with Thermo Scientific Imperial Protein Stain (Cat. No. 24615) and evaluated using Thermo Scientific myImageAnalysis Software (Cat. No. 62237) to determine purity. Total yield, recovery, and purity were similar for both resins.