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:

Bradford Dialysis 10 1.png

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