NanoBio:FPLC Methods

FPLC Methods

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CAjoFmethodHis

• Frac-950: serpentine row
• Variables:
  • Main:Column: HisTrap_FF_5mL
  • Start w/ Pump Wash:
    • Wash Inlet A: OFF
    • Wash Inlet B: OFF
  • Flow Rate: Flow Rate (mL/min): 0.7
  • Column Pressure Limit (MPa): 0.3
  • Averaging Time UV(s): 1.3
  • Eluent A Inlet: A2
  • Start Conc B (%): 0
  • Column Valve: Column_Position: Position 4
  • Column Equilibration: Equilibrate w/ (CV): 1.0
  • Aut_PressureFlow_Reg:
    • System Pump: Normal
    • System Press Level (MPa): 0
    • System Min Flow (mL/min): 0
  • Flowthrough Fractionation:
    • Tube Size (mm):18
    • Fraction Size (mL): 5
    • Flowthrough Fract Start at: First Tube
  • Sample Injection: Empty Loop w/ (mL): 2.0
  • Wash out Unbound Sample: Wash column w/ (CV): 10
  • Fractionation
    • Tube Type Eluate Fraction (mm): 18
    • Eluate Frac Size (mL): 1.0
    • Eluate Fract Start at: Next Tube
    • Tube Type Peak Fraction (mm): 18
    • Peak Frac Size (mL): 0
    • Peak Fract Start at: First Tube
  • Linear Gradient:
    • Target Conc B (%): 100
    • Length of Gradient (CV): 5
  • Gradient Delay: Gradient Delay (mL): 0
  • Clean after Elution: Clean w/ (CV): 2

CAjoFmethodHis1mL

• Note: same as CAjoFmethodHis except:
  • Main:Column: HisTrap_FF_1mL
  • Fractionation:Eluate Frac Size (mL): 0.25