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
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