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Experimental Biological Chemistry I
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Purpose
- To resuspend gold nano-particle pallets with Tris buffer.
- To determine the best concentration and pH of Tris buffer for resuspension via measuring the absorbance of resuspended solutions throughout time.
- To make new sets of Au/BSA solutions with varies ratios.
Procedure
- A new sets of solutions of BSA and HAuCl4 mixture were made. New stock solutions of HAuCl4 and BSA were also made.
- 100mM tris buffer at pH 8.0 and pH 10.0 were diluted with serial dilution to four different concentrations: 10mM, 1mM, 100uM, and 10uM, respectively.
- Resuspending Au/BSA solutions in Tris buffer
- The old sets of solutions of Au/BSA at different ratios were spin in centrifuge at 3000rpm for 5 minutes at room temperature.
- The supernatant were poured out. Pallet of different Au/BSA ratios were resuspended in different concentrations and different pH of Tris buffer.
- The details of buffer-pallet mix is shown in table below:
Ratio of Au/BSA Pallet
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Tris Buffer Concentrations [mM]
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Tris Buffer pH
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132
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10
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10
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133
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1
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10
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134
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0.1
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10
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136
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0.01
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10
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138
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10
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8
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140
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1
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8
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160
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0.1
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8
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170
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0.01
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8
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- The tris buffer and Au/BSA solution were mixed gently and let sit for 30 minutes.
- After 30 minutes, 500uL of each sample were loaded into quark cuvette and placed into UV-Vis to measure absorbance from 800nm to 200mm.
- Graph were plotted with Absorbance versus Wavelength, for details of absorbance from each sample, see Dhea Patel's Lab Notebook
Notes
- The new sets of solutions made on 2012/09/04 failed to react. The solutions after heating appeared yellow in color. This indicates the BSA failed to react with the HAuCl4 in solution.
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