User:Michael S. Bible/Notebook/571/2014/11/04

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Objective

  • Analyze dialysis solutions from yesterday
    • UV-Vis
    • Fluoresence
    • Bradford
    • Calcium ion ISE


  • Measure dialysis concentrations from last week
  • Prepare a new dialysis using colloid solution and CaCl2 or HCl
    • For those running CaCl2, use 3500 MWCO & test concentrations between 10 and 50 mM
    • For those running HCl, use 25000 MWCO & test concentrations between 100 μM and 10 mM
  • Prepare solutions for enzyme kinetics
    • Prepare 25 mL 66 mM Potassium phosphate, pH 6.24 (beaker prep is fine)
    • Place half in a Falcon tube, label & refrigerate overnight
    • Prepare 10 mL 0.01 wt% Micrococcus lysodeikticus Cell Suspension (in freezer) using phosphate buffer
    • Adjust the cell suspension concentration until the Absorbance at λ = 450 nm is between 0.6 & 0.7


Data

Ca2+ ISE for Lysozyme dialzyed against CaCl2

Original CaCl2 Concentration (mM) mV Measurement (mV) Final CaCl2 Concentration (mM)
6.83 55.0 2.68
1.87 39.9 0.684
0.68 2.9 0.024
0.020 -5.7 0.011
0.00058 -0.3 0.0179 (This is a bad data point)

New Dialysis of Colloid

Colloid was dialyzed against CaCl2 of the following concentrations

  • 45 mM
  • 35 mM
  • 25 mM
  • 15 mM
  • 5 mM