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|style="background-color: #EEE"|[[Image: | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab: Fall 2013</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> </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"|<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> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
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== | |||
* | ==Objective== | ||
To monitor the kinetics and yield of the [http://en.wikipedia.org/wiki/Horseradish_peroxidase horseradish peroxidase]-catalyzed oxidation of [http://en.wikipedia.org/wiki/Luminol luminol]. These experiments will be compared to future experiments with HRP-functionalized nanoparticles. These experiments are also meant to introduce researchers to stopped-flow techniques and rapid data collection. | |||
From [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/10/01 | Dr. Hartings]] | |||
==Procedure== | |||
'''2. Chemiluminescence of Luminol Oxidation Reaction Initiated by Stopped Flow Mixer''' | |||
# Add 64.9 μL HRP stock, 3.31 mL luminol stock, and 6.63 mL buffer to stopped flow mixer. | |||
# Add 5.00 mL hydrogen peroxide stock and 5.00 mL buffer to stopped flow mixer. | |||
# Equilibrate mixer tubes with sample. | |||
# Initiate mixing. | |||
# Measure light produced as result of reaction, integrated over a specific time range. | |||
# Integrate area under the curve. | |||
''' 3. Kinetics of Luminol Oxidized by Absorption Spectrum Changes in Stopped Flow Mixer''' | |||
# Add 245.13 μL HRP stock to a volumetric flask and dilute to 10 mL with buffer. | |||
# Add 1.25 mL luminol stock to a volumetric flask and dilute to 10 mL with buffer. | |||
# Add 2.00 mL of diluted HRP to stock and 10.00 mL of diluted luminol to stopped flow mixer. | |||
# Add 5.00 mL hydrogen peroxide stock and 5.00 mL buffer to stopped flow mixer. | |||
# Equilibrate mixer tubes with sample. | |||
# Initiate mixing. | |||
# Using luminol and 3-aminophthalic acid spectra as endpoints, determine the kinetics of 3-aminophthalic acid synthesis. | |||
==Figures== | |||
[[Image:KineticsUVVISluminol.jpg|700px|]] | |||
==Data== | |||
==Notes== | |||
* The stock solution of HRP was 0.77 μM. | |||
* The stock solution of luminol was 1.51 mM. | |||
* The buffer solution was 5.1 mM Tris at pH = 8. | |||
* The stock solution of hydrogen peroxide was 12.8 mM. | |||
Revision as of 09:19, 9 October 2013
Biomaterials Design Lab: Fall 2013 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectiveTo monitor the kinetics and yield of the horseradish peroxidase-catalyzed oxidation of luminol. These experiments will be compared to future experiments with HRP-functionalized nanoparticles. These experiments are also meant to introduce researchers to stopped-flow techniques and rapid data collection. From Dr. Hartings Procedure2. Chemiluminescence of Luminol Oxidation Reaction Initiated by Stopped Flow Mixer
FiguresDataNotes
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