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.
|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)
|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|
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.
For Research Use Only. Not for use in diagnostic procedures.