User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/28: Difference between revisions
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==Results for Running Au/ADA samples on UV-vis Spectrophotometer== | ==Results for Running Au/ADA samples on UV-vis Spectrophotometer== | ||
* The Au/ADA samples made with dialyzed ADA proteins all appeared purple with purple fiber formation at the bottom of tubes. A picture of the samples after 4 hours of heating is shown in picture below: | * The Au/ADA samples made with dialyzed ADA proteins all appeared purple with purple fiber formation at the bottom of tubes. A picture of the samples after 4 hours of heating is shown in picture below: | ||
[ | [[Image:Au ADA samples after dialysis.jpg|600px]] | ||
* The result indicated that the lack of salt in samples might have an affect on nanoparticle aggregation formation. The lack of salt in solution might lead to ADA proteins to wrap around gold in solution at a lower temperature. The formed gold nanoparticles might have different isoelectric points that engages protein aggregations. | * The result indicated that the lack of salt in samples might have an affect on nanoparticle aggregation formation. The lack of salt in solution might lead to ADA proteins to wrap around gold in solution at a lower temperature. The formed gold nanoparticles might have different isoelectric points that engages protein aggregations. | ||
* While the fiber formation can be due to lack of salt in solution that encouraged nanoparticle aggregation, the fiber formation might also be affected by the usage of plastic falcon tubes during the reaction. | * While the fiber formation can be due to lack of salt in solution that encouraged nanoparticle aggregation, the fiber formation might also be affected by the usage of plastic falcon tubes during the reaction. | ||
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* The data can be compared against the Au/ADA solution made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/27|2012/11/27]]. While the Au/ADA before dialysis appeared to have influence between the mole ratio of gold to ADA and concentration of gold nanoparticles in solution, the Au/ADA samples after dialysis does not. This can be explained by the presence of salt in solution that encouraged gold nanoparticle formation but can stabilize the protein nanoparticles to prevent nanoparticles from aggregating. | * The data can be compared against the Au/ADA solution made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/27|2012/11/27]]. While the Au/ADA before dialysis appeared to have influence between the mole ratio of gold to ADA and concentration of gold nanoparticles in solution, the Au/ADA samples after dialysis does not. This can be explained by the presence of salt in solution that encouraged gold nanoparticle formation but can stabilize the protein nanoparticles to prevent nanoparticles from aggregating. | ||
* Furthermore, the solutions for Au/ADA samples before dialysis did not appear purple, while solutions for Au/ADA samples after dialysis formed purple solutions and purple fibers. This can also be explained by the presence of salt in solution. The presence of salt might interact with gold nanoparticles in a way that encouraged gold nanoparticle configuration that does not absorbce at 525nm. The absence of salt in solution might allow gold nanoparticle to adapt another confirmation that does absorb at 525nm. | * Furthermore, the solutions for Au/ADA samples before dialysis did not appear purple, while solutions for Au/ADA samples after dialysis formed purple solutions and purple fibers. This can also be explained by the presence of salt in solution. The presence of salt might interact with gold nanoparticles in a way that encouraged gold nanoparticle configuration that does not absorbce at 525nm. The absence of salt in solution might allow gold nanoparticle to adapt another confirmation that does absorb at 525nm. | ||
==Procedure for Running Au/ADA samples on Atomic Absorption Spectrometer== | ==Procedure for Running Au/ADA samples on Atomic Absorption Spectrometer== | ||
* The same sample of Au/ADA were run on Atomic Absorption Spectrometer. | * The same sample of Au/ADA were run on Atomic Absorption Spectrometer. | ||
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==Results for Au/ADA Resuspension in Tris Buffer== | ==Results for Au/ADA Resuspension in Tris Buffer== | ||
* All solutions were successfully resuspended after the addition of Tris buffer. The solution after resuspension appeared as purple homogenous mixture and are clear in color. A picture taken after resuspension of all resuspended Au/ADA samples is shown below: | |||
[[Image:Au ADA after resuspension.jpg|600px]] | |||
* The color for solution for 110 Au/ADA were clearer than other Au/ADA samples because the solution was lighter in color before resuspension. | |||
* The amount of time it took for each solution to resuspend was recorded in table below: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Mole ratio of Au/ADA''' | |||
| align="center" style="background:#f0f0f0;"|'''Concentration of Tris buffer used[uM]''' | |||
| align="center" style="background:#f0f0f0;"|'''pH of Tris buffer''' | |||
| align="center" style="background:#f0f0f0;"|'''Time for resuspension[sec]''' | |||
|- | |||
| 60||1||10||3 | |||
|- | |||
| 70||10||10||4 | |||
|- | |||
| 80||100||10||4 | |||
|- | |||
| 90||1||8||7 | |||
|- | |||
| 100||10||8||6 | |||
|- | |||
| 110||100||8||6 | |||
|} | |||
* From table above, it can be concluded that resuspension happened faster in solutions with Tris buffer pH 10.0 then solutions with Tris buffer pH 8.0. | |||
* It was observed that resuspension time needed was must shorter in Tris buffer with pH 10.0 than in Tris buffer with pH 8.0. Possible reason for this result can be due to the high pH introduced by Tris buffer at pH 10.0 is much higher than the isoelectric point in ADA protein. This allows proteins to become negatively charged faster. Ions in buffer with stronger ionic strength can dissociate in solution and break apart the nanoparticle aggregation by interacting with the net negative charge on protein. | |||
* On the other side, at pH 8.0, less ADA proteins make the transition in time in order to take on a whole negative charge. As a result, not all ions would be able to form charge-charge interaction with the proteins, and in total delaying the process for protein aggregation breakdown, slowing down the time for complete resuspension. | |||
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Revision as of 21:11, 7 December 2012
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Purpose
Procedure for Running Au/ADA samples on UV-vis Spectrophotometer
60-70-80-90-100-110-120-130-140-150
Results for Running Au/ADA samples on UV-vis Spectrophotometer
Procedure for Running Au/ADA samples on Atomic Absorption Spectrometer
5-8-10-15-20-25-30-40
Results for Running Au/ADA samples on Atomic Absorption Spectrometer
Absorbance = 0.0153 * Concentration + 0.045
Procedure for Au/ADA Resuspension in Tris Buffer
Results for Au/ADA Resuspension in Tris Buffer
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