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

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Objectives

  1. Run fluorescence data for lysozyme at pH=4
  2. Create stock solutions for use on 09062016


Description

Fluorescence

  1. Solutions made using measurements below
  2. 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

  1. Solution Measurements
    1. Lysosome Stock Solution
      1. 8.88 mg of lysozyme with water in 10.0 mL volumetric flask
      2. MW = 14307 g/mol
      3. concentration = 62.1 μM
    2. Lysozyme: 0.25mM Au, 12.5μM Lysozyme, 3mL total; pH 4
      1. 225 μL AuCl3
      2. 300 μL 1M HCl
      3. 604 μL Lysozyme
      4. 1871 μL Water
  2. Fluorescence Summary Graphs
Fluorescence Emission 08312016 Lysozyme pH4.png

Figure 1 Maximum emission data of tryptophan in a solution of pH 4. Red points correspond to the times shown in Figure 3.

083116 Integ Fluor pH 4.png

Figure 2 Integrated fluorescence data of tryptophan in a solution of pH 4. Red points correspond to the times shown in Figure 3.

0831016 Full Spectra pH4.png

Figure 3 Emission summary of important points at t=0, 5, 35, 75, and 175.

Figure 3 Explanation

  1. At t=0 min, we see a max emission far to the left of the other emission spectra.
  2. At t=5 min, this maximum has shifted up and to the right.
  3. Until t=35 min, the maximum for each spectra decreases until the shown maximum.
  4. After t=35 min, the maximum begins increasing again.
  5. At t=75 min, the maximum continues to increase, but the "shoulder" seen at around 475 nm begins going away.
  6. 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

  1. AuCl3 stock A (from 8/30)
    1. mass = 10.13 mg
    2. 10.0 mL volumetric flask
    3. concentration = 3.34 mM
  2. AuCl3 stock B (from 8/31)
    1. mass = 12.6 mg
    2. 10.0 mL volumetric flask
    3. concentration = 4.17 mM
  3. Fructose (from 8/30)
    1. mass = 23.48 mg
    2. 10.0 mL volumetric flask
    3. concentration = 13.0 mM
  4. Lysozyme stock 1 (from 8/31)
    1. mass = 8.88 mg
    2. 10.0 mL volumetric flask
    3. concentration = 62.1 μM
  5. Lysozyme stock 2 (made today)
    1. mass = 21.92 mg
    2. 10.0 mL volumetric flask
    3. 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