User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/22

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Tasks for October 22

  • Data analysis for 30:1 colloid solution vs CaCl2 using MWCO 3500
  • New dialysis of 0.6 g/ L Lysozyme vs KI using 3500 MWCO since dialysis from Oct. 8 did not work due to incorrect dilutions

0.6g/L solution vs KI (MWCO 3500) Dialysis Data

Bradford Analysis

  • Only running bradford analysis of protein-containing solutions, that is Lys 1, 2, 3, 4 and 5, since the protein cannot diffuse through 3500 MWCO.
  • Remove 50 μL of solution from each chamber (7 in all) and run Bradford analysis
    • Bradford reagent should be diluted 1:3 with 50mM Tris/50mM NaCl
    • Recall, 50 μL solution + 200 μL diluted Bradford + 750 μL Tris/NaCl buffer
    • PS cuvettes, measuring 400 - 800 nm

Bradford assay 281014.png

I- ISE

I- concentration mV measurement (mV)
2mM (1) -175.3
Lys (1) -174.0
5 mM -196.4
Lys (2) -188.5
10 mM -208.9
Lys (3) -210.5
25 mM -231.5
Lys (4) -229.1
50 mM -250.2
Lys (5) -244.3

UV-Vis measurements

  • It was found that KI interferes with the Bradford reagent, so that no Bradford can bind to protein. No peak was seen using UV-Vis

Fluorescence measurements

  • No fluorescence measurements were taken on the samples, since nothing was detected using UV-Vis absorption.