User:Eleni N. Kalivas/Notebook/CHEM-571/2013/09/24: Difference between revisions
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==Objective== | |||
Analyze the catalytic activity of pepsin in the presence and absence of pepstatin. The data collected today will be compared to our data taken from pepsin-AuNPs. | |||
==Protocol== | ==Protocol== | ||
The following protocol was taken from [User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2013/09/24 Matt Hartings] | The following protocol was taken from [[User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2013/09/24|Matt Hartings]] | ||
We will allow the pepsin cleavage of hemoglobin to proceed for two hours with aliquots taken and analyzed every 1/2 hour. We will also run parallel reactions with different concentrations of pepstatin (pepsin inhibitor) present. We will save our aliquots for analysis tomorrow using SDS-PAGE. | We will allow the pepsin cleavage of hemoglobin to proceed for two hours with aliquots taken and analyzed every 1/2 hour. We will also run parallel reactions with different concentrations of pepstatin (pepsin inhibitor) present. We will save our aliquots for analysis tomorrow using SDS-PAGE. | ||
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### Measure the absorption spectrum (specifically note 280nm) in order to determine the protein concentration in solution. Use the stock hemoglobin solution as your reference | ### Measure the absorption spectrum (specifically note 280nm) in order to determine the protein concentration in solution. Use the stock hemoglobin solution as your reference | ||
# Repeat Step 3 every 30 minutes for 2 hours. | # Repeat Step 3 every 30 minutes for 2 hours. | ||
==UV vis== | |||
Absorbance Spectra of hemoglobin with pepsin | |||
[[Image:Pepsin09242013zem.png|750px]] | |||
Revision as of 10:28, 1 October 2013
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ObjectiveAnalyze the catalytic activity of pepsin in the presence and absence of pepstatin. The data collected today will be compared to our data taken from pepsin-AuNPs. ProtocolThe following protocol was taken from Matt Hartings We will allow the pepsin cleavage of hemoglobin to proceed for two hours with aliquots taken and analyzed every 1/2 hour. We will also run parallel reactions with different concentrations of pepstatin (pepsin inhibitor) present. We will save our aliquots for analysis tomorrow using SDS-PAGE.
UV visAbsorbance Spectra of hemoglobin with pepsin
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