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 PhyTip columns 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
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PhyNexus PhyTip GST-tagged Proteins
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.
Description

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.

Data

Shipping and Storage

Each pack of PhyTip columns has been manufactured and QC’d to the highest standards and shipped in retainer boxes that maintain the integrity of the specific affinity resin within each PhyTip column. This product is shipped at ambient temperatures, but on receipt should be stored in a standard laboratory refrigerator between 4 and 8◦C.

  • Do NOT freeze or store frozen.
  • When not in use, keep the lid of the box closed and sealed, store in the refrigerator.
  • Do not allow affinity resin to dry out by extended storage in a dry environment.

Glutathione PhyTip columns are stored in Glycerol when shipped from PhyNexus.

Documents

Glutathione Product Insert Sheet

Glutathione Flyer

Glutathione Spec Sheet

Glutathione PhyTip Columns MSDS

Robotics Systems Compatibility

GST-tagged Protein PhyTip Columns are available for these robotic systems.

Part Numbers

Glutathione PhyTip Columns