User:Carly M. Montanero/Notebook/CHEM-571/2013/10/02: Difference between revisions
From OpenWetWare
(Autocreate 2013/10/02 Entry for User:Carly_M._Montanero/Notebook/CHEM-571) |
(fix raw html notebook nav) |
||
(6 intermediate revisions by one other user not shown) | |||
Line 1: | Line 1: | ||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|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"| | |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]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | |||
* | ==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|]] | |||
==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. | |||
Latest revision as of 23:24, 26 September 2017
Biomaterials Design Lab: Fall 2013 | Main project page Previous entry Next entry |
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
Figures
Notes
|