User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2014/10/01

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Bradford Analysis of Lysozyme Dialysis

Bradford analysis of the dialysis was run for the soak solution and the protein solution. The results are below, along with calculated concentrations:

The soak solutions were not included in this graph, because the MW cutoff of the dialysis membrane prevented the Lysozyme from moving across the membrane.

Wavelength 575 Calculated Concentrations (g/L)
Stock0.1050.608040201
50 mM CaCl20.1080.75879397
50 mM CaCl2 Soak0.012-4.065326633
500 uM CaCl20.1030.507537688
500 uM CaCl2 Soak-0.009-5.120603015
HCl0.092-0.045226131
HCl Soak-0.005-4.91959799
Water0.1120.959798995
Water Soak-0.005-4.91959799


A second dialysis was prepared using 2 wells of 0.12 g/L lysozyme and the 30:1 Au:Lysozyme colloid against 20k MW paper. These were run against 50 mM, 500 µM, and 5 µM CaCl2. The dialysis will run for a week.

Ca2+ ISE

Water and the stock lysozyme solution (0.5 g/L) were tested with the Ca2+ ISE probe. The results are below:

Solution mV
Tap45.7
HPLC-78.5
DI-110.3
Lysozyme85.4



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