User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/27: Difference between revisions
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==Procedure for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples== | ==Procedure for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples== | ||
* UV-vis spectrometer were run in spectrum method. A range of 200nm to 800nm were ran on all Au/ADA and Au/HRP samples made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/14|2012/11/14]]. | * UV-vis spectrometer were run in spectrum method. A range of 200nm to 800nm were ran on all Au/ADA and Au/HRP samples both made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/14|2012/11/14]]. | ||
* 3mL of distilled water was placed in quartz cuvette with 1cm pathlength and set as base line solution. 3mL taken from supernatant of Au/ADA and Au/HRP samples were placed in an identical quartz cuvette with 1cm pathlength for spectra measurement. | * 3mL of distilled water was placed in quartz cuvette with 1cm pathlength and set as base line solution. 3mL taken from supernatant of Au/ADA and Au/HRP samples were placed in an identical quartz cuvette with 1cm pathlength for spectra measurement. | ||
* After measurement, each 3mL samples of Au/ADA and Au/HRP of different mole ratios were placed back into original sample test tubes to conserve products that would later be used for Atomic Absorption spectra analysis. | |||
==Results for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples== | ==Results for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples== | ||
* Au/ADA samples appeared clear, and no purple fibers were formed. This also indicates that no gold nanoparticles were present in solution. However, there were white precipitants observed at the bottom of the samples in samples with molar ratio 130 to 150. The white precipitants appear in small circular quantities and seemed to stick to the side of the glass test tube. A picture of white precipitant in Au/ADA with 150 mole ratio is shown below: | |||
[[Image:White precipitants.jpg|350px]] | |||
* It was suspected that the white precipitant could be precipitated ADA proteins when gold interacts with the salt in elution buffer. The elution buffer contained the following contents: 20mM Tris, 0.5M NaCl, and 500mM Imidazole at pH 7.5. Once salt is attracted to interact with HAuCl<sub>4</sub> in solution, ADA protein then precipitates due to lack of buffering for stabilization. | |||
* Au/HRP samples appeared purple in color. All samples had purple supernatant. Small fibers were observed for sample with mole ratio 130 to 150. For samples with 100 to 150 molar ratio, supernatant displayed a darker purple color compare to samples with 60 to 90 molar ratio. | |||
* UV-vis spectrometer were run for Au/ADA samples. Absorbance versus wavelength ranging from 200nm to 800nm were plotted on the graph titled "Absorbance of AuADA Solutions at Mole Ratios from 60 to 140 Au:ADA", shown below: | |||
[[Image:UV-Vis_Spectra_of_AuADA.JPG|600px]] | |||
* From graph above, it can be shown that Au/ADA samples have very low absorbance. This indicates that litle to none gold nanoparticles were formed within solution. It was also suspected a potential peak around 700nm from seeing the overall trend for all samples. However, that peak would most likely be due to the salt present in elution buffer. | |||
* UV-vis spectrometer were run for Au/HRP samples. Absorbance were graphed versus wavelength for samples with mole ratio ranging from 60 to 150. The graph titled "Absorbance of Solutions of AuHRP at Mole Ratios from 60 to 150 Au:HRP" is shown below: | |||
[[Image:UV-Vis_Spectra_of_AuADA_11142012.JPG|600px]] | |||
* From the graph, it can be suggested that 150 Au/HRP solution yielded the most amount of absorbance. This result seemed consistent to the procedure for making Au/HRP solutions since most amount of HAuCl<sub>4</sub> was placed into the 150 Au/HRP sample. | |||
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Revision as of 20:31, 27 November 2012
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Purpose
Procedure for Dialysis
Notes for Dialysis
Procedure for making dialyzed Au/ADA samples
60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150
Procedure for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples
Results for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples
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