User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/10/07

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Primary Experiment

Over the next 6 weeks, we will continue the dialysis experiments.
You will examine the following systems:

  • 0.6 g/L Lysozyme vs CaCl2 using MWCO 3500
  • 0.6 g/L Lysozyme vs HCl using MWCO 20000
  • 30:1 colloid solution vs CaCl2 using MWCO 3500
  • 30:1 colloid solution vs HCl using MWCO 20000
  • 0.6 g/L Lysozyme vs KI using MWCO 3500
  • 30:1 colloid solution vs KI using MWCO 3500

Measurements will include:

  • UV/VIS Bradford analysis (PS cuvettes) for all protein containing solutions
  • pH for all H+ containing solutions
  • Ca2+ ISE for all Ca2+ containing solutions
  • UV/VIS (glass microcuvette) for all protein solutions
  • Fluorescence (glass microcuvette & 100 fold diluted) for all protein solutions
  • Precipitation titration and/or fluorescence and/or electrochemistry for I- containing solutions

Consider the following as you decide which concentrations to test

  • Protein concentration in 30:1 colloid solution is about 8 μM
  • Prior class observed CaCl2 induced precipitation at 100 mM
  • Other scientists have found NaCl induced precipitation at 1.5 M
  • We want to determine if colloid precipitate occurs at same conditions as lysozyme precipitation
  • Lower concentrations are typically less precise

Additional Tasks During Waiting Periods

  • I- concentration using precipitation titration
  • I- concentration using potentiometer (Ag/AgI electrode)
  • Rerun gel electrophoresis
  • Test protein specific GPC column
  • Convert earlier GPC plots into Excel files
  • Lysozyme kinetics experiment

We will start with the I- concentration by titration today