Testing the effects of Tris buffer pH 10
- The prepared solutions of Au/BSA mole ratio 170 [1] are transferred from their glass test tubes to plastic centrifuge tubes.
- These centrifuge tubes containing the 170 Au/BSA solutions are placed inside the centrifuge (located in the balance room) under the conditions of 3000 rpm at 10°C for 5 min. It is relevant to centrifuge the solution to create a pellet of proteins that will separate it from its supernatant.
- Upon the termination of the centrifuge cycle, the supernatant was removed from the test tubes using plastic transfer pipets.
- The tris buffer pH 10 was taken from the refrigerator. Varying amounts of the buffer were distributed to the seven Au/BSA fibers by serial dilution. The distribution of buffer and water are listed in the table.
- When aliquot of buffers were done, the tubes were capped and vented to allow the solution to mix. UV-Vis scans were executed immediately. The first run of UV-Vis scans began at 1:30 PM. This is followed by two more runs which were executed at 1 h. intervals. Run 2 was started at 2:38 PM while run 3 was started at 3:35 PM.
- The UV-Vis scans of all tris buffer concentrations are shown below. Each graph show one particular concentration of tris buffer with three graphs pertaining to the three runs made in 1 h. intervals.
- Further procedures in exercising the effects of pH and ionic strength had been postponed to initiate the tasks of preparing for cellular growth and protein expression.
| Test Tube
|
volume of tris buffer pH 10 (mL)
|
volume of water (mL)
|
concentration of tris buffer (M)
|
| 1 |
10 |
0 |
.1
|
| 2 |
1 |
9 |
1x10-2
|
| 3 |
.1 |
9.9 |
1x10-3
|
| 4 |
.01 |
9.99 |
1x10-4
|
| 5 |
.001 |
9.999 |
1x10-5
|
| 6 |
.0001 |
9.9999 |
1x10-6
|
| 7 |
.00001 |
9.99999 |
1x10-7
|
Graphs of UV-Vis scans of Au/BSA in tris buffer pH 10
|
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