User:Moira M. Esson/Notebook/CHEM-571/2013/09/25: Difference between revisions
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#Run electrophoresis for 30 minutes at 200V | #Run electrophoresis for 30 minutes at 200V | ||
#Develop stain of the gel | #Develop stain of the gel | ||
#*Place gel in fixant solution (40% methanol, 10% acetic acid, 50% water) for | #*Place gel in fixant solution (40% methanol, 10% acetic acid, 50% water) for 20 minutes to stop the proteins from migrating further | ||
#*Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour | #*Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour | ||
#*Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes | #*Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes | ||
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Figure 1. SDS-PAGE Gel | |||
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[[Image:FLUBBER.jpg]] | |||
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==UV-vis== | |||
Figure 2. Corrected Absorbance Spectra of Pepsin with Hemoglobin | |||
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[[Image:Hemopepsin09251013zem.png]] | |||
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*Note: For the 2hr sample some pieces of protein may have been present when running the sample. | |||
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Figure 3. Corrected Absorbance Spectra of 20μM Pepstatin with Hemoglobin | |||
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[[Image:Pepstathemogl09252013zem.png]] | |||
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==Notes== | |||
*For both of the UV-vis spectra, there is no observable trend for the absorbance(the absorbance does not increase or decrease consistently with time). This may be due to the inclusion of protein in some of the sample runs. Another trial of these samples will be performed | |||
*Placed the gel in 10mLs of fixing solutions. Actually ended up dying the gel for 1hour and 15mins. | |||
Revision as of 17:34, 7 October 2013
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Objectives
Electrophoresis
UV-visFigure 2. Corrected Absorbance Spectra of Pepsin with Hemoglobin
Notes
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