High quality purification of different GST fusion proteins using Glutathione Agarose

Three GST-fusion proteins were expressed in E. coli, extracted in Thermo Scientific B-PER Reagent with Enzymes (Part No. 90078), and then purified using a Pierce Glutathione Agarose product. Elution fractions were separated on 4-20% gradient Tris-glycine gels and stained with Thermo Scientific GelCode Blue Stain Reagent (Part No. 24590).

Details:

  • Pak1-GST: Lysate (2.4mg total protein) was purified using the Pierce GST Spin Purification Kit (Part No.16106). Crude lysate was applied in Equilibration-Wash Buffer to a 0.2mL Pierce Glutathione Spin Column and then eluted with 10mM Glutathione Elution Buffer, pH 8.0. Yield of target Pak1-GST was 475µg.
  • Bid-GST: Expressing cells (225mg pellet) were lysed and the extract (11mg total protein) was applied to a 0.2mL Pierce Glutathione Spin Column (Part No. 16103) and eluted with 125mM Tris, 150mM NaCl, 10mM Glutathione, pH 8.0. Yield of target Bid-GST was 3.63mg.
  • ERK-GST: Expressing cells (480mg pellet) were lysed and 3mL cell extract (11.2 mg total protein) was applied in 50mM Tris, 150mM NaCl, pH 8.0 buffer to a column containing 0.5mL of Pierce Glutathione Agarose (Part No. 16100) and eluted with 125mM Tris, 150mM NaCl, 10mM Glutathione, pH 8.0. Yield of target Erk-GST was 1.132mg.

Three GST-fusion proteins were expressed in <em>E. coli</em>, extracted in Thermo Scientific B-PER Reagent with Enzymes (Part No. 90078), and then purified using a Pierce Glutathione Agarose product. Elution fractions were separated on 4-20% gradient Tris-glycine gels and stained with Thermo Scientific GelCode Blue Stain Reagent (Part No. 24590).</p><h3>Details:</h3><ul><li><strong>Pak1-GST:</strong> Lysate (2.4mg total protein) was purified using the Pierce GST Spin Purification Kit (Part No.16106). Crude lysate was applied in Equilibration-Wash Buffer to a 0.2mL Pierce Glutathione Spin Column and then eluted with 10mM Glutathione Elution Buffer, pH 8.0. Yield of target Pak1-GST was 475µg.</li><li><strong>Bid-GST:</strong> Expressing cells (225mg pellet) were lysed and the extract (11mg total protein) was applied to a 0.2mL Pierce Glutathione Spin Column (Part No. 16103) and eluted with 125mM Tris, 150mM NaCl, 10mM Glutathione, pH 8.0. Yield of target Bid-GST was 3.63mg.</li><li><strong>ERK-GST:</strong> Expressing cells (480mg pellet) were lysed and 3mL cell extract (11.2 mg total protein) was applied in 50mM Tris, 150mM NaCl, pH 8.0 buffer to a column containing 0.5mL of Pierce Glutathione Agarose (Part No. 16100) and eluted with 125mM Tris, 150mM NaCl, 10mM Glutathione, pH 8.0. Yield of target Erk-GST was 1.132mg.</li></ul>

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