User:Sara K. Mattson/Notebook/Chemistry 671/2016/09/06

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Objective

Florescence Spectroscopy is used to determine the rate at which myoglobin unfolds at pH of 4.

Myoglobin Fluoresence pH 4

  1. Stock soltions used:
    1. 5.08mM Au
    2. 13.0mM Fructose
    3. 183uM Myoglobin
  2. Myoglobin: 0.381mM Au, 28.304uM Myoglobin, 3mL total
    1. 225uL AuCl3
    2. 300uL 1M HCl
    3. 464uL Myoglobin
    4. 1871uL Water

Description

  1. Absorbance measurements were taken and saved every 5 minutes for 3 hours
  2. The instrument was set at 80 degrees Celsius
  3. The scan range parameters were start at 310nm and ending at 540nm
  4. The emission and exit slit were set at 10nm

Results

Image:Myoglobin wavelength ph4.png Image:Integrated intensity myglobin ph4.png Image:First 15 minutes myoglobin ph4.png Image:35-45 minutes myoglobin ph4.png

Solutions Made for UV- VIS Tomorrow

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 Myoglobinstock μL of Water
4 500 0 0 500 0 342 3658
5 50 0 0 500 0 342 4108
6 5 0 0 500 0 342 4153
7 0 0 0 500 0 342 4158
8 0 5 0 500 0 342 4153
9 0 50 0 500 0 342 4108
10 0 500 0 500 0 342 3658
11 0 0 5 500 0 342 4153
12 0 0 50 500 0 342 4108
0.625 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 Myoglobinstock μL of Water
4 500 0 0 500 240 342 3418
5 50 0 0 500 240 342 3868
6 5 0 0 500 240 342 3913
7 0 0 0 500 240 342 3918
8 0 5 0 500 240 342 3913
9 0 50 0 500 240 342 3868
10 0 500 0 500 240 342 3418
11 0 0 5 500 240 342 3913
12 0 0 50 500 240 342 3868

Notes

The concentration of fructose in the second chart was made at 0.625mM as opposed to 0.0625mM. The concentration of Myoglobin was 28.304uM as opposed to 12.5uM and the concentration of Au was 0.381mM as opposed to 0.225mM. All of the solutions made were a light orange color except for 0.625mM fructose at pH 12, which was a light purple color when made.


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