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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PhyNexus PhyTip GST-tagged Proteins

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.

PhyNexus Dual Flow Chromatography PhyNexus PhyTip columns used with our proprietary Dual Flow Chromatography provide complete antibody binding for highly concentrated sample recovery from small samples and can be used in all major 8, 12 or 96 column robotic liquid handling systems.
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