User:Cosette Brianna Taggart/Notebook/Experimental/2016/08/31

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August 31, 2016 : Fluorescence I

Objectives

  1. Collect fluorescence data for lysozyme at pH 4
  2. Make stock solutions for UV-Vis on 9.06.2016

Description

Fluoresence

  1. Lysozyme solution was made according to the measurements found in data below
  2. Lysozyme solution was run through florescence for 3 hours with data taken every 5 minutes

Stock Solutions for 9.06.2016

Solutions were made in glass test tubes, covered with foil, and placed in an oven for 4 hours at 80 C.

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


SUMMARY DATA AND FIGURES

Summary Data at 15 minute increments
Time(minutes) Maximum Emission Integrated Intensity
0 360.5 7641.99525
15 427 9442.508657
30 427 8162.719866
45 425 8304.910077
60 431 8762.672858
75 433 9335.867079
90 431 9865.379939
105 432.5 20446.13203
120 431 21129.30793
135 426 21739.42892
150 429.5 22169.51961
165 426 22522.05809
180 430 14677.52683

MaxEM083116.png

IntegratedIntensity083116.png

Points of Interest

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.

Integrated Fluorescence 083116 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.

Integration Summary

Integration Summary 83116 Lysozyme pH 4.png

Figure 3 Full emission spectra for selected time points of Figure 1 and Figure 2

  • Matt Hartings 11:47, 6 September 2016 (EDT): do add these when you get the chance.

Solution Prep

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


UPDATE

Below are corrected solution prep 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 14:04, 1 September 2016 (EDT): Your sample prep didn't work. See my lab notebook page here for a description of why and how I remade your samples.
  • Matt Hartings 12:52, 2 September 2016 (EDT): I have taken the complementary data for a sample with no gold. Please see data here