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
|
Tris Buffer Concentrations [mM]
|
Tris Buffer pH
|
| 132
|
10
|
10
|
| 133
|
1
|
10
|
| 134
|
0.1
|
10
|
| 136
|
0.01
|
10
|
| 138
|
10
|
8
|
| 140
|
1
|
8
|
| 160
|
0.1
|
8
|
| 170
|
0.01
|
8
|
- 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.
|
Experimental Biological Chemistry I
