User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/11: Difference between revisions

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==Procedure==
==Procedure==
* Two samples, one at 120 Au/BSA and another at 60 Au/BSA were made with stock solutions. The two samples were placed in incubator for an hour to check for possible reaction.
* Two samples, one at 120 Au/BSA and another at 60 Au/BSA were made with stock solutions at concentrations used for 2012/09/04. The two samples were placed in incubator for an hour to check for possible reaction.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:52, 17 September 2012 (EDT)''':what stock solution? concentrations? when were they prepared? volumes and masses?
* UV absorbance were taken from samples that were mixed with Tris buffer from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05|2012/09/05]]. Sample at the ratio 134, 136, 138, 140, 160, 170 Au to BSA were centrifuged at 4000rpm, 7°C, for 5 minutes to ensure separation of supernatant and pallet. 500uL of each supernatant were used in each quartz cuvette. Each sample were run from 800nm to 200nm. The graph Absorbance versus Wavelength were plotted and shown in [[User:Melissa Novy/Notebook/CHEM-571/2012/09/11|Melissa's Notebook]].
* UV absorbance were taken from samples that were mixed with Tris buffer from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05|2012/09/05]]. Sample at the ratio 134, 136, 138, 140, 160, 170 Au to BSA were centrifuged at 4000rpm, 7°C, for 5 minutes to ensure separation of supernatant and pallet. 500uL of each supernatant were used in each quartz cuvette. Each sample were run from 800nm to 200nm. The graph Absorbance versus Wavelength were plotted and shown in [[User:Melissa Novy/Notebook/CHEM-571/2012/09/11|Melissa's Notebook]].
* After an hour in the incubator, the 120 Au/BSA ratio solution turned to transparent, and the purple fibers formed in 60 Au/BSA ratio solutions. New sets of HAuCl4 and BSA solutions at different ratios were made with below amounts:
* After an hour in the incubator, the 120 Au/BSA ratio solution turned to transparent, and the purple fibers formed in 60 Au/BSA ratio solutions. New sets of HAuCl4 and BSA solutions at different ratios were made with below amounts:
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* Sets of sample of Au/BSA with different ratios appeared gold in color after four hours in incubation. This indicates HAuCl4 has failed to react with BSA in solution.
* Sets of sample of Au/BSA with different ratios appeared gold in color after four hours in incubation. This indicates HAuCl4 has failed to react with BSA in solution.
* Au/BSA solutions mixed in tris buffer had a pellet at the bottom of the 50mL falcon tube, supernatent were carefully taken. For some samples the pellets were disturbed when transferring. To obtain pure supernatent, the falcon tubes were centrifuged.
* Au/BSA solutions mixed in tris buffer had a pellet at the bottom of the 50mL falcon tube, supernatent were carefully taken. For some samples the pellets were disturbed when transferring. To obtain pure supernatent, the falcon tubes were centrifuged.
 
* From the absorbance measurements, we concluded that all Au/BSA ratios worked successfully, results appeared identical to the results obtained from past experiments. All ratios for Au/BSA were concluded to be correct.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:53, 17 September 2012 (EDT)''':it is unclear what conclusions you drew from the absorbance measurements?




Revision as of 08:58, 5 October 2012

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Purpose

  • Check if newly made sample on 2012/09/05 has reacted after 4 hours in the incubator at 85°C
  • Measure UV absorbance on samples that were diluted in Tris buffer in different concentration and pH

Procedure

  • Two samples, one at 120 Au/BSA and another at 60 Au/BSA were made with stock solutions at concentrations used for 2012/09/04. The two samples were placed in incubator for an hour to check for possible reaction.
  • UV absorbance were taken from samples that were mixed with Tris buffer from 2012/09/05. Sample at the ratio 134, 136, 138, 140, 160, 170 Au to BSA were centrifuged at 4000rpm, 7°C, for 5 minutes to ensure separation of supernatant and pallet. 500uL of each supernatant were used in each quartz cuvette. Each sample were run from 800nm to 200nm. The graph Absorbance versus Wavelength were plotted and shown in Melissa's Notebook.
  • After an hour in the incubator, the 120 Au/BSA ratio solution turned to transparent, and the purple fibers formed in 60 Au/BSA ratio solutions. New sets of HAuCl4 and BSA solutions at different ratios were made with below amounts:
HAuCl4/BSA ratio'' BSA[uL]'' HAuCl4[uL]'' H2O[mL]''
60 165 32.2 3.803
80 165 43.0 3.792
100 165 53.7 3.781
120 165 64.5 3.771
128 165 68.7 3.766
130 165 69.8 3.765
132 165 70.9 3.764
133 165 71.4 3.764
134 165 72.0 3.763
136 165 73.0 3.762
138 165 74.1 3.761
140 165 75.2 3.760
160 165 85.9 3.749
170 165 91.3 3.744

Notes

  • Sets of sample of Au/BSA with different ratios appeared gold in color after four hours in incubation. This indicates HAuCl4 has failed to react with BSA in solution.
  • Au/BSA solutions mixed in tris buffer had a pellet at the bottom of the 50mL falcon tube, supernatent were carefully taken. For some samples the pellets were disturbed when transferring. To obtain pure supernatent, the falcon tubes were centrifuged.
  • From the absorbance measurements, we concluded that all Au/BSA ratios worked successfully, results appeared identical to the results obtained from past experiments. All ratios for Au/BSA were concluded to be correct.



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