Do I need to equilibrate the column before use?

Yes, the column will need to be equilibrated before use. Ensure that the columns are at room temperature with both caps on, before operation. Operational temperature is 18 - 24°C.

  • If an elution buffer other than PBS is to be used, equilibrate the column with at least 3x the column volume of elution buffer.
  • Do not allow the column to run dry. The frit must stay wet. If the column runs dry it may not function correctly.
  • Use only fresh filtered (0.22 μm) buffer to avoid introducing particulate contamination.
  • For convenience and best results purchase Izon Reagent Kits for TRPS use. 
  • Results may be erroneous if operated outside the operation temperature range.

    qEVsingle Gen 2 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the buffer volume.
    2. Once column is at room temperature, leave the bottom luer-slip cap in place. 
    3. Remove the top cap carefully and replace the electrolyte in the top of the column with elution buffer
    4. Remove the lower luer-slip cap and rinse the column with at least 6 mL of elution buffer (we use PBS electrolyte). If an elution buffer other than PBS is to be used, equilibrate the column with at least 9 mL of elution buffer. 
    5. Note the time for 5 mL of buffer to flow through – this is useful for detecting when to clean the column. 
    6. Monitor the top of the frit. Do not let the column run dry at any point.

    qEVoriginal Gen 2 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the buffer volume.
    2. Once column is at room temperature, leave the bottom luer-slip cap in place. 
    3. Remove the top cap carefully and replace the electrolyte in the top of the column with elution buffer
    4. Remove the lower luer-slip cap and rinse the column with at least 17 mL of elution buffer (we use PBS electrolyte). If an elution buffer other than PBS is to be used, equilibrate the column with at least 25.5 mL of elution buffer. 
    5. Note the time for 5 mL of buffer to flow through – this is useful for detecting when to clean the column. 
    6. Monitor the top of the frit. Do not let the column run dry at any point.

     

    qEV2 Gen 2 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the buffer volume.
    2. Once column is at room temperature, leave the bottom end cap in place.
    3. Rinse the reservoir with electrolyte.
    4. Unscrew the connector cap carefully and fill in the connector with filtered electrolyte - try to avoid getting air bubbles into the column or running the column dry. 
    5. Screw in the reservoir and load 30 mL of filtered electrolyte in the reservoir. 
      • Flush through with a total of 90 mL of buffer. If an elution buffer other than PBS is to be used, equilibrate the column with at least 135 mL of elution buffer.
    6. Remove the bottom cap and let the electrolyte run through.
    7. Note the time for 5 mL of buffer to flow through – this is useful for detecting when to clean the column, if the flow rate drops by 25%, the column should be cleaned. 

     

    qEV10 Gen 2 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the buffer volume.
    2. Once column is at room temperature, leave the bottom end cap in place.
    3. Rinse the reservoir with electrolyte.
    4. Unscrew the connector cap carefully and fill in the connector with filtered electrolyte - try to avoid getting air bubbles into the column or running the column dry. 
    5. Screw in the reservoir and load 60 mL of filtered electrolyte in the reservoir. 
      • Flush through with a total of 140 mL of buffer. If an elution buffer other than PBS is to be used, equilibrate the column with at least 210 mL of elution buffer.
    6. Remove the bottom cap and let the electrolyte run through.
    7. Note the time for 5 mL of buffer to flow through – this is useful for detecting when to clean the column, if the flow rate drops by 25%, the column should be cleaned.