User:Eleni N. Kalivas/Notebook/CHEM-571/2013/10/01: Difference between revisions
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== | ==Objective== | ||
==Protocol== | |||
the following protocol was taken from [[User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2013/10/01|Matt Hartings]] | |||
Three sets of measurements will be performed today. | |||
# UV-Vis Absorbance of reactants, catalysts, and products | |||
## horseradish peroxidase (Use stock solution) | |||
## luminol (use stock solution) | |||
## 3-aminophthalic acid (product of reaction between luminol and H2O2 catalyzed by HRP. In order to take this measurement, react (in 1:1 ratio or with slight excess H2O2) luminol with H2O2 in presence of HRP. Allow the reaction to proceed for 5 minutes; take the spectrum) | |||
# Chemiluminescence of luminol oxidation reaction initiated by stopped flow mixer | |||
## Add HRP/Luminol stock solution to stopped flow mixer | |||
## Add H2O2 stock solution 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 | |||
# Kinetics of luminol oxidized by changes in absorption spectrum, reaction carried out in stopped flow mixer | |||
## Add HRP/Luminol stock solution to stopped flow mixer | |||
## Add H2O2 stock solution 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. | |||
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ObjectiveProtocolthe following protocol was taken from Matt Hartings Three sets of measurements will be performed today.
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