Objectives
AA spectroscopy
General Protocol:
- Transfer ~4mL of sample from test tubes into a 10mL falcon tube.
- Centrifuge ~4mL protein-AuNP solutions for 30 minutes at 4000rpm. Be sure that the centrifuge is balanced.
- Prepare 5 standard Au solutions (10μg/mL, 20μg/mL, 30μg/mL, 40μg/mL, 50μg/mL) following protocol described on 09/10/2013.
- Run samples on AA.
- Prepare calibration curve and prepare a graph showing the concentration of Au vs Au:Protein ratio
Table 1. Standard Au Solution preparation from 1000μg/mL AA Au gold standard solution
| Concentration(μg/mL)
|
Volume of stock Au solution(mL)
|
Volume Deionized water added(mL)
|
| 10 |
0.1 |
9.9
|
| 20 |
0.2 |
9.8
|
| 30 |
0.3 |
9.7
|
| 40 |
0.4 |
9.6
|
| 50 |
0.5 |
9.5
|
Figure 1. AA Gold Standard Calibration Curve
Figure 2. Hemoglobin Absorbance Curve
Figure 3. Myoglobin Absorbance Curve
Notes
- Lysozyme solutions prepared on 10/30/2013 all produced fibers and appear colorless except for the 30:1 ratio(appears furthest to the right). This solution is mildly colored.
- All myoglobin solutions prepared on 10/30/2013 produced fibers and appear colorless.
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Project name
