User:Hannah Harvey/Notebook/CHEM-471 Fall 2016/2016/08/31

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Objectives Run fluorescence data for lysozyme at pH=4 Create stock solutions for use on 09062016 Description Fluorescence Solutions made using measurements below Fluorescence data run every 5 minutes for 3 hours Solutions for 09/06/2016 Solutions were added to glass test tubes, covered with foil, and placed in an oven set to 80°C for 4 hours. Data Fluorescence Solution Measurements Lysosome Stock Solution 8.88 mg of lysozyme with water in 10.0 mL volumetric flask MW = 14307 g/mol concentration = 62.1 μM Lysozyme: 0.25mM Au, 12.5μM Lysozyme, 3mL total; pH 4 225 μL AuCl3 300 μL 1M HCl 604 μL Lysozyme 1871 μL Water Fluorescence Summary Graphs Figure 1 Maximum emission data of tryptophan in a solution of pH 4. Red points correspond to the times shown in Figure 3. Figure 2 Integrated fluorescence data of tryptophan in a solution of pH 4. Red points correspond to the times shown in Figure 3. Figure 3 Emission summary of important points at t=0, 5, 35, 75, and 175. 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=75 min, the maximum continues to increase, but the "shoulder" seen at around 475 nm begins going away. t=175 shows the final full spectra obtained. Solution Prep These were the initial solutions made with the wrong concentration of lysozyme. See the last 2 tables for the correct solutions. Lysozyme UVVis solution measurement 0 mM fructose pH HCl (μL) NaOHc (μL) NaOHd (μL) AuCl3 (μL) Fructose (μL) Lysozyme (μL) Water (μL) 4 500 0 0 300 0 252 3949 5 50 0 0 300 0 252 4399 6 5 0 0 300 0 252 4444 7 0 0 0 300 0 252 4449 8 0 0 5 300 0 252 4444 9 0 0 50 300 0 252 4399 10 0 0 500 300 0 252 3949 11 0 5 0 300 0 252 4444 12 0 50 0 300 0 252 4399 Lysozyme UVVis solution measurement .0625 mM fructose pH HCl (μL) NaOHc (μL) NaOHd (μL) AuCl3 (μL) Fructose (μL) Lysozyme (μL) Water (μL) 4 500 0 0 300 24 252 3925 5 50 0 0 300 24 252 4375 6 5 0 0 300 24 252 4420 7 0 0 0 300 24 252 4425 8 0 0 5 300 24 252 4420 9 0 0 50 300 24 252 4375 10 0 0 500 300 24 252 3925 11 0 5 0 300 24 252 4420 12 0 50 0 300 24 252 4375 Below are the correct solutions re-made by Dr. Hartings stock solutions AuCl3 stock A (from 8/30) mass = 10.13 mg 10.0 mL volumetric flask concentration = 3.34 mM AuCl3 stock B (from 8/31) mass = 12.6 mg 10.0 mL volumetric flask concentration = 4.17 mM Fructose (from 8/30) mass = 23.48 mg 10.0 mL volumetric flask concentration = 13.0 mM Lysozyme stock 1 (from 8/31) mass = 8.88 mg 10.0 mL volumetric flask concentration = 62.1 μM Lysozyme stock 2 (made today) mass = 21.92 mg 10.0 mL volumetric flask concentration = 153 μM 0 mM Fructose pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water 4 500 0 0 300B 0 10061 3194 5 50 0 0 300B 0 10061 3644 6 5 0 0 300B 0 10061 3689 7 0 0 0 300B 0 10061 3694 8 0 5 0 300B 0 4082 4287 9 0 50 0 300B 0 4082 4242 10 0 500 0 300B 0 4082 3792 11 0 0 5 300B 0 4082 4287 12 0 0 50 300B 0 4082 4242 0.0625 mM Fructose pH μL of 1 mM HClstock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AuCl3stock μL of Fructosestock μL of BSAstock μL of Water 4 500 0 0 300B 24 4082 3768 5 50 0 0 300B 24 4082 4218 6 5 0 0 300B 24 4082 4263 7 0 0 0 300B 24 4082 4268 8 0 5 0 300B 24 4082 4263 9 0 50 0 374A 24 4082 4144 10 0 500 0 374A 24 4082 3694 11 0 0 5 374A 24 4082 4189 12 0 0 50 374A 24 4082 4144 Matt Hartings 12:52, 2 September 2016 (EDT): I have taken the complementary data for a sample with no gold. Please see data here