User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2014/10/14: 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>}}
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| align="center" style="background:#f0f0f0;"|'''Wavelength'''
| align="center" style="background:#f0f0f0;"|'''Wavelength'''
| align="center" style="background:#f0f0f0;"|'''577'''
| align="center" style="background:#f0f0f0;"|''' Abs at 577 nm'''
| align="center" style="background:#f0f0f0;"|'''Concentration (ug/mL)'''
| align="center" style="background:#f0f0f0;"|'''Concentration (ug/mL)'''
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'''Fluorescence'''
[[Image:Lysozyme_KI_3500_Fluorescence_Emission_Chart.png|500px]]
'''UV-VIS'''
[[Image:Lysozyme_KI_3500MW_UV_VIS_1in100_Chart.png|500px]]





Latest revision as of 00:26, 27 September 2017

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SDS-PAGE Prep

SDS-PAGE was prepared according to the following:

  • 10 μL 0.6 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
  • 10 μL 0.12 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
  • 10 μL 30:1 Au/lysozyme colloid with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
  • 10 μL 0.12 g/L unknown protein with 10 μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
  • Placed in heating block (set at 90 °C) for 5 minutes, and placed in the fridge overnight.


A pH 6.24 66 mM Potassium Phosphate buffer was prepared using monobasic and dibasic solids.


Dialysis Analysis

Bradford, UV-VIS, and Fluorescence analysis were run on the dialysis samples from the KI dialysis. 1 in 4 Bradford solution was prepared and the samples were prepped the same method as previously. A blank was also run. For UV-VIS and fluorescence, a 1 in 100 dilution of each was prepared and run. UV-VIS was run from 200-400 nm.


Bradford Results

Wavelength Abs at 577 nm Concentration (ug/mL)
Stock 0.121 1.169902913
2 mM 0.115 0.878640777
5 mK 0.124 1.315533981
10 mM 0.114 0.830097087
25 mM 0.111 0.684466019
50 mM 0.115 0.878640777

Fluorescence

UV-VIS