User:Sara K. Mattson/Notebook/Chemistry 671/2016/09/27

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Fluorescence

Purpose

Use fluorescence to examine the rate at which myoglobin unfolds at pH4

Procedure

  1. Sample was made Myoglobin: 0.25mM Au, 12.5uM Myoglobin, 1.0mM HCl 3mL total
    1. Au 4.57mM
      1. (0.25mM)(3.0mL)=(XmL)(4.57mM)
      2. XmL= .176mL= 176uL Au
    2. Myoglobin 160uM
      1. (12.5uM)(3.0mL)=(XmL)(160uM)
      2. X=0.234mL= 234uL Myoglobin
    3. HCl 1.0mM
      1. (3.0mL)(1.0mM)=(1.0mM)(XmL)
      2. XmL=3.0mL= 300uL HCl
    4. Water
      1. Volume of water= 3000uL- 176uL-234uL-300uL= 2290uL
  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

Solutions for UV-VIS preparation

Stock Concentrations:

  1. Au 4.57mM
  2. Myoglobin 160mM
  3. Fructose 40.9mM
0.25 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 0 293 31 388 3788
5 50 0 0 293 31 388 4238
6 5 0 0 293 31 388 4283
7 0 0 0 293 31 388 4288
8 0 5 0 293 31 388 4283
9 0 50 0 293 31 388 4238
10 0 500 0 293 31 388 3788
11 0 0 5 293 31 388 4283
12 0 0 50 293 31 388 4238
0.75 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 0 293 92 388 3727
5 50 0 0 293 92 388 4177
6 5 0 0 293 92 388 4222
7 0 0 0 293 92 388 4227
8 0 5 0 293 92 388 4222
9 0 50 0 293 92 388 4177
10 0 500 0 293 92 388 3727
11 0 0 5 293 92 388 4222
12 0 0 50 293 92 388 4177


1.25 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 0 293 159 388 3666
5 50 0 0 293 159 388 4116
6 5 0 0 293 159 388 4161
7 0 0 0 293 159 388 4166
8 0 5 0 293 159 388 4161
9 0 50 0 293 159 388 4116
10 0 500 0 293 159 388 3666
11 0 0 5 293 159 388 4161
12 0 0 50 293 159 388 4116

Results

After the sample was run through fluorescence, no color change had occurred indicating there was a source of error in the data collected, possibly from there being no gold in the sample prepared. The graphs for the incorrect data were still made and are shown below. The correct sample was run on 09/29/2016 by Matt Hartings and the correct graphs were created.

Graphs For Incorrect Data

Integrated intensity for Myoglobin templated Gold Nanoparticles at pH4

Integrated intensity gold nanoparticles pH4 myoglobin.png

Maximum intensity for Myoglobin templated Gold Nanoparticles at pH4

Max intensity for gold nanoparticle myoglbin at ph 4.png

Maximum Wavelength for Myoglobin templated Gold Nanoparticles at pH4

Maxium wavelength myoglobin gold ph4.png


  1. The graphs for the correct data were created by Matt Hartings. Data was collected and interpreted on 09/28/2016

Notes

pH4 was already done on 08/31/2016 but the prepared solution had incorrect concentrations of myoglobin and gold NPs