User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/13 - Revision history

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 08 Dec 2012 04:54:14 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 04:54, 8 December 2012
Line 106:		Line 106:
	* 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.
-	* ~~Please refer to [[User:Melissa Novy/Notebook/CHEM-571/2012/11/13\|Melissa Novy's Notebook]] for graph~~ and data plotting the percent concentration change before and after in unit of ppm for both Au/HRP and Au/Lysozyme.	+	* 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==		==Procedure on making adenosine==

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 08 Dec 2012 04:25:30 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 04:25, 8 December 2012
Line 106:		Line 106:
	* 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.
		+	* Please refer to [[User:Melissa Novy/Notebook/CHEM-571/2012/11/13\|Melissa Novy's Notebook]] for graph and data plotting the percent concentration change before and after in unit of ppm for both Au/HRP and Au/Lysozyme.

	==Procedure on making adenosine==		==Procedure on making adenosine==

Keyun Wang: /* Procedure on making adenosine */

Keyun Wang — Sat, 24 Nov 2012 17:05:28 GMT

Procedure on making adenosine

←Older revision		Revision as of 17:05, 24 November 2012
Line 108:		Line 108:

	==Procedure on making adenosine==		==Procedure on making adenosine==
-	* Adenosine was made to prepare for ADA activity assay.	+	* 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.		* 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.		* The pH of sodium phosphate buffer was adjusted to 7.4 using HCl.

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:35:08 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:35, 24 November 2012
Line 97:		Line 97:
	\| 450\|\|25.6198\|\|130\|\|-2.6454		\| 450\|\|25.6198\|\|130\|\|-2.6454
	\|}		\|}
		+
	* 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]]
Line 104:		Line 105:
	[[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.
		+
	==Procedure on making adenosine==		==Procedure on making adenosine==
	* Adenosine was made to prepare for ADA activity assay.		* Adenosine was made to prepare for ADA activity assay.

Keyun Wang: /* Procedure on making adenosine */

Keyun Wang — Sat, 24 Nov 2012 03:34:14 GMT

Procedure on making adenosine

←Older revision		Revision as of 03:34, 24 November 2012
Line 110:		Line 110:
	* The pH of sodium phosphate buffer was adjusted to 7.4 using HCl.		* 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.		* 0.1mM of adenosine was made in 5mL of sodium phosphate buffer.
-	* 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.	+	* 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.
-	* This procedure was not done by writer, thus was not described in details.	+
	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
	\|}		\|}

	__NOTOC__		__NOTOC__

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:33:41 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:33, 24 November 2012
Line 105:		Line 105:
	* 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.
		+	==Procedure on making adenosine==
		+	* Adenosine 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.
		+	* 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.
		+	* This procedure was not done by writer, thus was not described in details.
	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
	\|}		\|}

	__NOTOC__		__NOTOC__

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:27:08 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:27, 24 November 2012
Line 103:		Line 103:
	* Gold concentrations in Au/Lysozyme samples were also determined and plotted onto the graph "Concentration of Au in Solution of Au-Lysozyme" below:		* Gold concentrations in Au/Lysozyme samples were also determined and plotted onto the graph "Concentration of Au in Solution of Au-Lysozyme" below:
	[[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.
		+	* 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.
	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
	\|}		\|}

	__NOTOC__		__NOTOC__

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:22:30 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:22, 24 November 2012
Line 103:		Line 103:
	* Gold concentrations in Au/Lysozyme samples were also determined and plotted onto the graph "Concentration of Au in Solution of Au-Lysozyme" below:		* Gold concentrations in Au/Lysozyme samples were also determined and plotted onto the graph "Concentration of Au in Solution of Au-Lysozyme" below:
	[[Image:AAS_AuLys.JPG\|600px]]		[[Image:AAS_AuLys.JPG\|600px]]
-	*	+	*
	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
	\|}		\|}

	__NOTOC__		__NOTOC__

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:21:44 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:21, 24 November 2012
Line 104:		Line 104:
	[[Image:AAS_AuLys.JPG\|600px]]		[[Image:AAS_AuLys.JPG\|600px]]
	*		*
		+	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
		+	\|}
		+
		+	__NOTOC__

Keyun Wang: /* Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions */

Keyun Wang — Sat, 24 Nov 2012 03:20:38 GMT

Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions

←Older revision		Revision as of 03:20, 24 November 2012
Line 69:		Line 69:

	==Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions==		==Data on Atomic Absorption Spectrometry of Au/ADA and Au/HRP solutions==
-	~~<!-- ##### DO NOT edit below this line unless you know what you are doing~~. ##### -->	+	* The concentration of gold in unit of ppm was calculated by converting absorbance into concentration using the HCl calibration curve.
		+	* The concentration of gold for corresponding ratios of Au/HRP and Au/Lysozyme were displayed in table below:
		+	{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|'''Au/HRP ratio'''
		+	\| align="center" style="background:#f0f0f0;"\|'''Concentration of Au[ppm]'''
		+	\| align="center" style="background:#f0f0f0;"\|'''Au/Lysozyme Ratio'''
		+	\| align="center" style="background:#f0f0f0;"\|'''Concentration of Au[ppm]'''
		+	\|-
		+	\| 10\|\|-1.5443\|\|20\|\|0.0057
		+	\|-
		+	\| 50\|\|1.8191\|\|30\|\|1.9388
		+	\|-
		+	\| 100\|\|11.7714\|\|40\|\|2.7586
		+	\|-
		+	\| 150\|\|13.1419\|\|50\|\|3.806
		+	\|-
		+	\| 200\|\|23.6748\|\|60\|\|4.5002
		+	\|-
		+	\| 250\|\|24.4588\|\|70\|\|-2.4599
		+	\|-
		+	\| 300\|\|27.876\|\|80\|\|-2.4839
		+	\|-
		+	\| 350\|\|25.4761\|\|100\|\|-2.5856
		+	\|-
		+	\| 400\|\|35.1173\|\|120\|\|-2.6394
		+	\|-
		+	\| 450\|\|25.6198\|\|130\|\|-2.6454
	\|}		\|}
-		+	* A graph was made with concentration of gold in ppm in supernatant versus mole ratio of Au to HRP and displayed below:
-	~~__NOTOC__~~	+	[[Image:AAS_AuHRP.JPG\|600px]]
		+	* From the graph above, increase in mole ratio of Au/HRP leads to an linear trend of increase in gold concentration in supernatant. This indicates no protein aggregation would form as mole ratio of Au/HRP increases. The result provide a contrast with results from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/17\|2012/10/17]] with the maximum concentration of gold in supernatant at low mole ratios of Au/BSA and minimum concentration of gold in supernatant once the mole ratios of Au/BSA reaches over 134.
		+	* The result indicate that protein aggregation is less likely to form when HRP is used to replace BSA. This might be caused by the nature of HRP compare to BSA and different behaviors when proteins unfold and refold.
		+	* Gold concentrations in Au/Lysozyme samples were also determined and plotted onto the graph "Concentration of Au in Solution of Au-Lysozyme" below:
		+	[[Image:AAS_AuLys.JPG\|600px]]
		+	*