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)
Stock 0.105 0.608040201
50 mM CaCl2 0.108 0.75879397
50 mM CaCl2 Soak 0.012 -4.065326633
500 uM CaCl2 0.103 0.507537688
500 uM CaCl2 Soak -0.009 -5.120603015
HCl 0.092 -0.045226131
HCl Soak -0.005 -4.91959799
Water 0.112 0.959798995
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
Tap 45.7
HPLC -78.5
DI -110.3
Lysozyme 85.4



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