User:Megan L. Channell/Notebook/Horseradish/2013/10/23

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Objective

  1. To take the spectra of the pepsin-AuNP samples made yesterday
  2. To make new HRP and Myoglobin samples

Description

  1. Protein-AuNP analysis
    1. Take a picture and note physical description of each ratio
    2. Place 1mL of each ratio in an 1.5mL eppendorf tube.
    3. Centrifuge for 30 minutes (Be considerate of other uses here. Act quickly, and get as many tubes in the centrifuge as you can at once.)
    4. Take a UV-Vis spectrum of each sample (Be considerate of other users here)
    5. After taking the spectrum, save the sample in a fresh eppy tube.
  2. Synthesis of HRP-AuNPs and Hemoglobin-AuNPs
    1. Into test tube that will contain a total of 5mL add
    2. Au solution such that the final concentration is 0.25mM
    3. For Myoglobin make the following solutions to reflect an Au:Protein ratios
      1. 30
      2. 50
      3. 70
      4. 90
      5. 110
      6. 130
      7. 150
      8. 170
      9. 190
      10. 210
    4. For HRP make the following solutions to reflect an Au:Protein ratios
      1. 230
      2. 250
      3. 270
      4. 290
      5. 310
      6. 330
      7. 350
      8. 370
      9. 390
      10. 410
    5. Water up to 5mL

Data

  • How solutions were made:

CHEM571 10.23.13 solutionstable.png

  • HRP UV-Vis

11.1.2013 cmj UVVis HRPhigh.png

  • Myoglobin UV-Vis

11.1.2013 cmj UVVis myo.png

Notes

Stock Solutions

  1. Gold
    1. 49.3 mg HAuCl4·3H2O in 50mL water ---> 2.50mM
  2. HRP
    1. 6.6 mg HRP in 100 mL water ---> 1.5μM
  3. Myoglobin
    1. 28.7mg myoglobin in 100mL water ---> 16.9μM