User:Monika Gasiorek/Notebook/CHEM-571 2014F/2014/11/05

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November 5, 2014 Primary Experiment #5 Work-up 30:1 colloid solution vs KI using MWCO 3500 Well 1 Well 2 Well 3 Well 4 Well 5 30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid 2 mM KI 5 mM KI 10 mM KI 25 mM KI 50 mM KI Necessary Analyses: Bradford 5 colloid samples, 30:1 colloid reference, and Bradford blank 200 μL sample, 200 μL 1:4 Bradford, 600 μL 50 mM Tris/50 mM NaCl Results will be posted Fluorescence & UV-VIS not necessary per NOTE on 10/21 I- ISE 10 samples Concentration Colloid Solution Potential (eV) Soak Solution Potential (eV) 50 mM -243.0 -245.2 25 mM -225.2 -222.5 10 mM -202.8 -204.9 5 mM -186.9 -187.9 2 mM -163.0 -165.1 Primary Experiment #6 Set-up 0.6 g/L Lysozyme vs CaCl2 using MWCO 3500 Well 1 Well 2 Well 3 Well 4 Well 5 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozome 45 mM CaCl2 35 mM CaCl2 25 mM CaCl2 15 mM CaCl2 5 mM CaCl2 NOTE: Due to the results of previous experiments, it has been observed that the lysozyme (and colloid) precipitate out of solution at concentrations between 25 and 50 mM. Additionally, Ca2+ ISE measurements were unstable at concentrations below 0.5 mM. The Ca2+ concentration range has been adjusted accordingly. Enzyme Kinetics Kinetics mode on UV-VIS 10mL of 400 unit/mL of lysozyme was prepared Cold buffer was used The procedure in the protocol was followed (see link in yesterday's tasklist) The protocol was repeated using 4 solutions between 200 & 400 units/mL Preparation was done just before addition to keep solution cold NOTE: We never actually got to do this because the UV was not accessible...