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 30 mL of elution buffer for 10 mL columns and at least 10 mL of elution buffer for 3 mL columns.
  • 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 qNano use. 
  • Results may be erroneous if operated outside the operation temperature range.

     

    qEVoriginal and qEVsingle columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the void 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 10 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 30 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 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the void 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 fill in 20-30 mL of filtered electrolyte in the reservoir. 
      • If an elution buffer other than PBS is to be used, equilibrate the column with at least 120 mL of elution buffer.
    6. Remove the bottom cap and let the electrolyte run through.
    7. Note the time for 2 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 columns

    1. Place the column in a holder and level it (make sure the column is vertical). Prepare a large vessel to collect the void 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 fill in 40-50 mL of filtered electrolyte in the reservoir. 
      • If an elution buffer other than PBS is to be used, equilibrate the column with at least 250 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.