- Run fluorescence data for lysozyme at pH=5
- Create stock solutions for use on 09.14.2016
- Solutions made using measurements below
- Fluorescence data run every 5 minutes for 3 hours
Solutions for 09/14/2016
Solutions were added to glass test tubes, covered with foil, and placed in an oven set to 80°C for 4 hours.
- Solution Measurements
- Lysosome Stock Solution
- 33.32 mg of lysozyme with 10.0 mL of water
- MW = 14307 g/mol
- concentration = 233 μM
- Gold Stock Solution
- 34.62 mg of AuCl3 with 25.0 mL of water
- concentration = 4,57 mM
- Lysozyme: 0.25mM Au, 12.5μM Lysozyme, 3mL total; pH 5
- 164 μL AuCl3
- 30 μL 1M HCl
- 161 μL Lysozyme
- 2645 μL Water
- Fluorescence Summary Graphs
Figure 1 Maximum emission data of tryptophan in a solution of pH 5.
Figure 2 Integrated fluorescence data of tryptophan in a solution of pH 5.
Figure 3 Full emission spectra for important time points: t=0, 5, 35, 90, 180 min.
Figure 3 Explanation
- At t=0 min, we see a max emission far to the left of the other emission spectra.
- At t=5 min, this maximum has shifted up and to the right.
- Until t=35 min, the maximum for each spectra decreases until the shown maximum.
- After t=35 min, the maximum begins increasing again.
- At t=90 min, the maximum continues to increase, but the "shoulder" seen at around 475 nm begins going away.
- t=180 shows the final full spectra obtained.
UV-vis solutions for 09.14.2016
Table 1 UV-vis solutions made for [Fructose]=.125mM
Table 2 UV-vis solutions made for [Fructose]=.25mM
At t=0, the cuvette had been in the machine briefly for about 1-2 minutes before the machine was started, which could account for some error in the data.
While running the fluorescence data, the machine briefly shut off at the 100 minute mark. The fluorescence data shows t=95, t=103, t=105 instead.
After taking the UV-vis samples out of the 80°C oven, pH 11 for both Fructose concentrations showed a precipitate form. pH 12 did not show this precipitate.