Fluorescence
Purpose
Use fluorescence to examine the rate at which Myoglobin templated gold nanoparticles unfold at pH6
Procedure
- Sample was made Myoglobin: 0.25mM Au, 12.5uM Myoglobin, .001mM HCl 3mL total
- Au 2.26mM
- (0.25mM)(3.0mL)=(XmL)(2.26mM)
- XmL= .332mL= 332uL Au
- Myoglobin 160uM
- (12.5uM)(3.0mL)=(XmL)(160uM)
- X=0.234mL= 234uL Myoglobin
- HCl 0.001mM
- (3.0mL)(0.001mM)=(1.0mM)(XmL)
- XmL=0.003mL= 3uL HCl
- Water
- Volume of water= 3000uL- 332uL-234uL-3uL= 2431uL
- Absorbance measurements were taken and saved every 3 minutes for 3 hours
- The instrument was set at 80 degrees Celsius
- The scan range parameters were start at 310nm and ending at 540nm
- The emission and exit slit were set at 10nm
Graphs
Wavelength of Myoglobin AuNp's at pH6
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Maximum Intensity of Myoglobin AuNp's at pH6
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Integrated Intensity of Myoglobins AuNp's at pH6
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Solution for UV-Vis Preparation
Stock Concentrations:
- Myoglobin 190uM
- Silver 16.6mM
- 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.
|