Glutathione Columns

Glutathione PhyTip Columns demonstrate high binding affinity and high capacity for the affinity purification of functional recombinant GST-tagged proteins.

  • Glutathione columns routinely produce samples of greater than 90% purity.
  • 60-80% recovery of sample proteins.
  • Glutathione tips achieve high levels of purity even in the presence of 40X excess BSA
  • High yields are obtained regardless of the level of contaminant proteins in the sample
  • Note that the proper use of Glutathione PhyTip columns requires slow flow rates of 0.06mL/min

Glutathione columns with our proprietary Dual Flow Chromatography provide complete antibody binding and high final concentrations. PhyTip columns are compatible with most 8, 12, and 96-channel automation liquid handling robots including Agilent Technologies, Beckman Coulter, Dynamic Devices, Hamilton, Opentrons, Perkin Elmer, and Tecan.

Related: IMAC (His-tagged Proteins)  Streptavidin (Biotin-tagged Molecules)

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Glutathione-GST-tagged-Proteins-PhyTip-Columns

Glutathione tips are available in tip volumes (µL) of 200|300|500|1000 and resin volumes (µL) of 5|10|20|40|80|160|320 depending upon robot configuration.

SDS-PAGE gel of GST enriched from varying levels of background proteins using Glutathione PhyTip columns. Purified protein was eluted from the column with 20 mM reduced glutathione. Lane 1: MW ladder Lane 2: 2 µg pure GST Lanes 3,4: Duplicate enrichments of a 25 µg/mL sample of GST from PBSwith 1 mg/mL BSA Lanes 5,6: Duplicate enrichments of a 25 µg/mL sample of GST from PBS.
PhyNexus Dual Flow Chromatography

Glutathione PhyTip columns have been optimized for use with specific PhyNexus reagents and instrument flow rates/volumes as shown below. This information was collected using the PhyTip ME 1000 and ME 200 Purification Systems.

NOTE THAT FLOW RATE IS OF CRITICAL IMPORTANCE WHEN PERFORMING GLUTATHIONE-BASED SEPARATIONS; SLOWER FLOW RATES ARE TYPICALLY REQUIRED AS COMPARED TO OTHER RESINS!

200+ PhyTip columns with Protein A resin:

A 200 µL E. coli lysate sample with 6 µg GST was processed using the conditions shown below. Greater than 20% of the original GST mass is recovered in the final sample volume. In addition, this recovered GST is purified to >95% purity as determined by HPLC with UV absorbance detection at 215 nm.

Capture: 200 µL sample is processed by passing through the resin bed for two cycles at 60 uL/minute.

Purify: 200 µL of PhyNexus Glutathione Wash Buffer I is passed through the resin bed for two cycles at 500 µL/min, followed by a second wash with the same buffer passed through the resin bed for two cycles at 500 µL/min.

Enrich: elute the protein into solution with 10µL of PhyNexus Glutathione Elution Buffer passed through the resin bed for four cycles at a flow rate of 500 µL/min.

1000+ PhyTip columns with Protein A resin:

A 500 µL E. coli lysate sample with 15 µg GST was processed using the conditions shown below. Greater than 20% of the original GST mass is recovered in the final sample volume. In addition, this recovered GST is purified to >95% purity as determined by HPLC with UV absorbance detection at 215 nm.

Capture: 500 µL sample is processed by passing through the resin bed for two cycles at 60 uL/minute.

Purify: 1000 µL of PhyNexus Glutathione Wash Buffer I is passed through the resin bed for two cycles at 500 µL/min, followed by a second wash with the same buffer passed through the resin bed for two cycles at 500 µL/min.

Enrich: elute the protein into solution with 15µL of PhyNexus Glutathione Elution Buffer passed through the resin bed for four cycles at a flow rate of 500 µL/min.

Select your resin, robotic system, tip and resin volume and see the results in the table below.

Glutathione Columns