User:Sara K. Mattson/Notebook/Chemistry 671/2016/10/05

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Fluorescence

Purpose

Use fluorescence to examine the rate at which Myoglobin templated gold nanoparticles unfold at pH6

Procedure

  1. Sample was made Myoglobin: 0.25mM Au, 12.5uM Myoglobin, .001mM HCl 3mL total
    1. Au 2.26mM
      1. (0.25mM)(3.0mL)=(XmL)(2.26mM)
      2. XmL= .332mL= 332uL Au
    2. Myoglobin 160uM
      1. (12.5uM)(3.0mL)=(XmL)(160uM)
      2. X=0.234mL= 234uL Myoglobin
    3. HCl 0.001mM
      1. (3.0mL)(0.001mM)=(1.0mM)(XmL)
      2. XmL=0.003mL= 3uL HCl
    4. Water
      1. Volume of water= 3000uL- 332uL-234uL-3uL= 2431uL
  2. Absorbance measurements were taken and saved every 3 minutes for 3 hours
  3. The instrument was set at 80 degrees Celsius
  4. The scan range parameters were start at 310nm and ending at 540nm
  5. The emission and exit slit were set at 10nm

Graphs

Wavelength of Myoglobin AuNp's at pH6

Maximum Intensity of Myoglobin AuNp's at pH6

Integrated Intensity of Myoglobins AuNp's at pH6

Solution for UV-Vis Preparation

Stock Concentrations:

  1. Myoglobin 190uM
  1. Silver 16.6mM
  1. Fructose 40.9mM


0 mM Fructose
pH μL of 1 mM HNO3stock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AgNO33stock μL of Fructosestock μL of Myoglobinstock μL of Water
4 500 0 0 75 0 329 4096
5 50 0 0 75 0 329 4546
6 5 0 0 75 0 329 4591
7 0 0 0 75 0 329 4596
8 0 5 0 75 0 329 4591
9 0 50 0 75 0 329 4546
10 0 500 0 75 0 329 4091
11 0 0 5 75 0 329 4591
12 0 0 50 75 0 329 4546


0.0625 mM Fructose
pH μL of 1 mM HNO3stock μL of 1 mM NaOHstock μL of 1 M NaOHstock μL of AgNO33stock μL of Fructosestock μL of Myoglobinstock μL of Water
4 500 0 0 75 8 329 4088
5 50 0 0 75 8 329 4538
6 5 0 0 75 8 329 4583
7 0 0 0 75 8 329 4588
8 0 5 0 75 8 329 4583
9 0 50 0 75 8 329 4538
10 0 500 0 75 8 329 4088
11 0 0 5 75 8 329 4583
12 0 0 50 75 8 329 4538

Notes

The silver nitrate needed to be handled in a dark room, and the test tubes were completely wrapped in tin foil because it is a light sensitive solution.



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