User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/11/28

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==Objective==
==Objective==
-
*to analyze the AuADA mixtures from [[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/11/27|yesterday]] using UV-spectroscopy and AAS.
+
*to analyze the AuADA mixtures from [[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/11/27|yesterday]] using UV-spectroscopy and AAS, and
 +
*to resuspend Au/ADA in Tris Buffer.
==Description==
==Description==
 +
 +
''UV-vis analysis of AuADA post-dialysis solutions''
#3mL samples of AuADA mixtures ranging form molar ratios of 60 to 150 Au:ADA were run in the UV-vis.  
#3mL samples of AuADA mixtures ranging form molar ratios of 60 to 150 Au:ADA were run in the UV-vis.  
#The data was collected, corrected for water, and plotted.
#The data was collected, corrected for water, and plotted.
 +
 +
''AAS analysis of AuADA post-dialysis solutions''
 +
#HCl was used as a blank and the AAS was auto-corrected for water.
 +
#8 standards were used: 5ppm Au, 10ppm Au, 15pp Au, 20ppm Au, 25ppm Au, 30ppm Au, 35ppm Au, and 40ppm Au.
 +
#Each sample of AuHRP and AuADA was run and the measured concentration was collected and plotted.
 +
 +
''Please refer to [[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/11/28| Kay's entry]] for discussion on the resuspension of AuADA in Tris Buffer.''
 +
 +
==Data/Observations==
 +
 +
[[Image:Au_ADA_samples_after_dialysis.jpg]]
 +
*This is an image of the Au/ADA samples made with dialyzed ADA protein. After the reaction, all the samples contained purple fibers at the bottom of the tubes.
 +
*A likely reason for the difference between the solutions formed with pre-dialyzed ADA (which produced clear samples) and the dialyzed ADA was the lack of imidazole salt in the ADA's solution.
 +
 +
[[Image:UV-Vis_Spectra_of_AuADA.JPG]]
 +
*This graph shows no significant difference in Au concentration in the samples, as the absorbance values were within 0.005 AU of each other.
 +
 +
[[Image:Water_and_Au_ADA.jpg]]
 +
*This graph shows also depicts the patter seen in the previous graph.
 +
 +
'''AAS'''
 +
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Au/ADA mole ratios'''
 +
| align="center" style="background:#f0f0f0;"|'''Absorbance(Abs.)'''
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| align="center" style="background:#f0f0f0;"|'''Concentration[ppm]'''
 +
|-
 +
| 60||0.0657||1.3572
 +
|-
 +
| 70||0.041||-0.2608
 +
|-
 +
| 80||0.1011||3.6762
 +
|-
 +
| 90||0.0292||-1.0337
 +
|-
 +
| 100||0.0246||-1.331
 +
|-
 +
| 110||0.0259||-1.2499
 +
|-
 +
| 120||0.0246||-1.3351
 +
|-
 +
| 130||0.0222||-1.4923
 +
|-
 +
| 140||0.0264||-1.2172
 +
|-
 +
| 150||0.0229||-1.4464
 +
|}
 +
*This table show the absorbance and concentration values of the AuADA solutions.
 +
 +
[[Image:Concentration versus Au ADA mole ratios.png|550px]]
 +
*This graph shows the concentration of Au in ppm vs the Au/ADA samples in mole ratios.
 +
*The peak at 80 Au:ADA mole ratio was due to an accidental absorbance of Au fibers.
 +
*The graph shows that the Au concentration in Au/ADA samples remain relatively the same through out the various Au:ADA mole ratio solutions.
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Objective

  • to analyze the AuADA mixtures from yesterday using UV-spectroscopy and AAS, and
  • to resuspend Au/ADA in Tris Buffer.

Description

UV-vis analysis of AuADA post-dialysis solutions

  1. 3mL samples of AuADA mixtures ranging form molar ratios of 60 to 150 Au:ADA were run in the UV-vis.
  2. The data was collected, corrected for water, and plotted.

AAS analysis of AuADA post-dialysis solutions

  1. HCl was used as a blank and the AAS was auto-corrected for water.
  2. 8 standards were used: 5ppm Au, 10ppm Au, 15pp Au, 20ppm Au, 25ppm Au, 30ppm Au, 35ppm Au, and 40ppm Au.
  3. Each sample of AuHRP and AuADA was run and the measured concentration was collected and plotted.

Please refer to Kay's entry for discussion on the resuspension of AuADA in Tris Buffer.

Data/Observations

Image:Au_ADA_samples_after_dialysis.jpg

  • This is an image of the Au/ADA samples made with dialyzed ADA protein. After the reaction, all the samples contained purple fibers at the bottom of the tubes.
  • A likely reason for the difference between the solutions formed with pre-dialyzed ADA (which produced clear samples) and the dialyzed ADA was the lack of imidazole salt in the ADA's solution.

Image:UV-Vis_Spectra_of_AuADA.JPG

  • This graph shows no significant difference in Au concentration in the samples, as the absorbance values were within 0.005 AU of each other.

Image:Water_and_Au_ADA.jpg

  • This graph shows also depicts the patter seen in the previous graph.

AAS

Au/ADA mole ratios Absorbance(Abs.) Concentration[ppm]
600.06571.3572
700.041-0.2608
800.10113.6762
900.0292-1.0337
1000.0246-1.331
1100.0259-1.2499
1200.0246-1.3351
1300.0222-1.4923
1400.0264-1.2172
1500.0229-1.4464
  • This table show the absorbance and concentration values of the AuADA solutions.

  • This graph shows the concentration of Au in ppm vs the Au/ADA samples in mole ratios.
  • The peak at 80 Au:ADA mole ratio was due to an accidental absorbance of Au fibers.
  • The graph shows that the Au concentration in Au/ADA samples remain relatively the same through out the various Au:ADA mole ratio solutions.


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