User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/13: Difference between revisions
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* A graph was made with concentration of gold in ppm in supernatant versus mole ratio of Au to HRP and displayed below: | * A graph was made with concentration of gold in ppm in supernatant versus mole ratio of Au to HRP and displayed below: | ||
[[Image:AAS_AuHRP.JPG|600px]] | [[Image:AAS_AuHRP.JPG|600px]] | ||
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[[Image:AAS_AuLys.JPG|600px]] | [[Image:AAS_AuLys.JPG|600px]] | ||
* In the graph above, the concentration of gold in Au/Lysozyme supernatant increased from mole ratio of 20 to 60, and dropped immediately from mole ratio of 60 and on. From mole ratio of 60 and on, there are no gold present in supernatant. The samples with mole ratio from 60 and on appear transparent as oppose to purple. | * In the graph above, the concentration of gold in Au/Lysozyme supernatant increased from mole ratio of 20 to 60, and dropped immediately from mole ratio of 60 and on. From mole ratio of 60 and on, there are no gold present in supernatant. The samples with mole ratio from 60 and on appear transparent as oppose to purple. | ||
* This result indicates that protein started to form aggregation from mole ratio of 60 and on. Compare to the result with Au/HRP from above and Au/BSA from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/17|2012/10/17]], the behavior of Au/Lysozyme resemble more like Au/BSA samples in terms of protein aggregation formation. This result also indicates that BSA and lysozyme behavior similarly when forming gold nano particles. It can also indicate that they have similar protein folding and unfolding kinetic mechanism. | * This result indicates that protein started to form aggregation from mole ratio of 60 and on. Compare to the result with Au/HRP from above and Au/BSA from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/17|2012/10/17]], the behavior of Au/Lysozyme resemble more like Au/BSA samples in terms of protein aggregation formation. This result also indicates that BSA and lysozyme behavior similarly when forming gold nano particles. It can also indicate that they have similar protein folding and unfolding kinetic mechanism. | ||
* Graph and data plotting the percent concentration change before and after in unit of ppm for both Au/HRP and Au/Lysozyme is shown below: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Au/ADA ratio''' | |||
| align="center" style="background:#f0f0f0;"|'''ADA added [uL]''' | |||
| align="center" style="background:#f0f0f0;"|'''HAuCl4 [uL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Water [uL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Mol Au [mol]''' | |||
| align="center" style="background:#f0f0f0;"|'''Mass Au [mg]''' | |||
| align="center" style="background:#f0f0f0;"|'''Initial [Au] [ppm]''' | |||
| align="center" style="background:#f0f0f0;"|'''Final [Au] [ppm]''' | |||
| align="center" style="background:#f0f0f0;"|'''Change in [Au] [ppm]''' | |||
| align="center" style="background:#f0f0f0;"|'''Percent Change [Au] [%]''' | |||
|- | |||
| 60||137.1||45.71||7817.2||4.79955E-07||0.094535296||11.81691206||1.3572||10.45971206||88.51476601 | |||
|- | |||
| 70||137.1||53.3||7809.6||5.5965E-07||0.110232582||13.77907269||-0.2608||14.03987269||101.8927253 | |||
|- | |||
| 80||137.1||60.9||7802||6.3945E-07||0.125950548||15.74381852||3.6762||12.06761852||76.64988328 | |||
|- | |||
| 90||137.1||68.6||7794.4||7.203E-07||0.14187533||17.73441626||-1.0337||18.76811626||105.8287794 | |||
|- | |||
| 100||137.1||76.2||7786.8||8.001E-07||0.157593297||19.69916209||-1.331||21.03016209||106.7566326 | |||
|- | |||
| 110||137.1||83.8||7779.1||8.799E-07||0.173311263||21.66390791||-1.2499||22.91380791||105.7695038 | |||
|- | |||
| 120||137.1||91.4||7771.5||9.597E-07||0.18902923||23.62865374||-1.3351||24.96375374||105.6503431 | |||
|- | |||
| 130||137.1||99||7763.9||1.0395E-06||0.204747197||25.59339956||-1.4923||27.08569956||105.8308002 | |||
|- | |||
| 140||137.1||106.7||7756.3||1.12035E-06||0.220671978||27.58399731||-1.2172||28.80119731||104.4127034 | |||
|- | |||
| 150||137.1||114.3||7748.7||1.20015E-06||0.236389945||29.54874313||-1.4464||30.99514313||104.8949629 | |||
|} | |||
* The table above listed the calculated initial gold concentration in unit of ppm and final measured concentration of gold in ppm in Au/ADA dialyzed samples. The change in gold concentration indicates how much gold nanoparticles were formed. If the percent change of gold is creater than 1, then it means that most of gold were incorporated into gold nanoparticles. When percent change goes below 1, data indicates that there were still some percentage of gold left in solution that were not nucleated within proteins to for gold nanoparticles. | |||
* The graph plotting the percent change in concentration of gold in solution versus the mole ratios of Au/ADA were shown in graph below: | |||
[[Image:AuADA_AAS_Percent.jpg]] | |||
* It can be seen that th percent change in gold concentration were relatively the same. The lowest percent change in gold concentration appeared to be 80Au/ADA. However, this might be due to accidental suspension of nanoparticle fibers when Atomic absorption spectra were taken. | |||
* Aside from the 80Au/ADA data point, the second lowest change in percent change in gold concentration happened to be 60 Au/ADA. This result might indicate that the concentration of gold in solution in 60 folds encouraged less nanoparticle formation compared to Au/ADA samples with greater mole ratios. | |||
* Lastly, the percent chage in gold concentration were roughly the same in remaining Au/ADA samples as the mole ratios goes above 90. This indicates that as the mole ratio increase, more gold were incorporated within gold nanoparticles. This indicates that high mole ratio in solution help facilitate gold nanoparticle nucleation. | |||
==Procedure on making adenosine== | |||
* Adenosine free base was obtained in powder form from aMR-ESCO®. | |||
* Sodium phosphate dibasic Heptahydrate was obtained from Fisher Scientific®. | |||
* Diluted adenosine in liquid form was made to prepare for ADA activity assay. | |||
* 0.05M sodium phosphate buffer was made in 50mL of distilled water. | |||
* The pH of sodium phosphate buffer was adjusted to 7.4 using HCl. | |||
* 0.1mM of adenosine was made in 5mL of sodium phosphate buffer. | |||
* This procedure was not done by writer, thus was not described in details. For details of the calculations, see [[User:Melissa Novy/Notebook/CHEM-571/2012/11/13|Melissa's Notebook]], which includes calculations for making sodium phosphate buffer, pH adjustment of sodium phosphate buffer, calculations to weight out the adenosine, and calculations on the amount of sodium phosphate buffer to add to adenosine in order to reach 0.1mM adenosine. | |||
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Revision as of 21:54, 7 December 2012
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Purpose
Procedure on UV-vis spectrometry of Au/HRP
10 - 50 - 100 - 150 - 200 - 250 - 300 - 350 - 400 - 450
Data on UV-vis spectrometry of Au/HRP
Procedure on Atomic Absorption Spectrometry of Au/lysozyme and Au/HRP solutions
5ppm - 8ppm - 10ppm - 15ppm - 20ppm - 25ppm - 30ppm - 40ppm
Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions
Procedure on making adenosine
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