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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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Bradford analysis of the dialysis was run for the soak solution and the protein solution. The results are below, along with calculated concentrations:
Bradford analysis of the dialysis was run for the soak solution and the protein solution. The results are below, along with calculated concentrations:


[[Image:Bradford_Analysis_Dialysis_10_1.png|500px]]
[[Image:Bradford_Dialysis_10_1.png|500px]]


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.


{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Wavelength'''
| align="center" style="background:#f0f0f0;"|'''575'''
| align="center" style="background:#f0f0f0;"|'''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 CaCl<sub>2</sub>. The dialysis will run for a week.
==Ca<sup>2+</sup> ISE==
Water and the stock lysozyme solution (0.5 g/L) were tested with the Ca<sup>2+</sup> ISE probe. The results are below:
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Solution'''
| align="center" style="background:#f0f0f0;"|'''mV'''
|-
| Tap||45.7
|-
| HPLC||-78.5
|-
| DI||-110.3
|-
| Lysozyme||85.4
|}




Latest revision as of 00:22, 27 September 2017

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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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