Objective
- To take the spectra of the pepsin-AuNP samples made yesterday
- To make new HRP and Myoglobin samples
Procedure
- Protein-AuNP analysis
- Take a picture and note physical description of each ratio
- Place 1mL of each ratio in an 1.5mL eppendorf tube.
- 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.)
- Take a UV-Vis spectrum of each sample (Be considerate of other users here)
- After taking the spectrum, save the sample in a fresh eppy tube.
- Synthesis of HRP-AuNPs and Hemoglobin-AuNPs
- Into test tube that will contain a total of 5mL add
- Au solution such that the final concentration is 0.25mM
- For Myoglobin make the following solutions to reflect an Au:Protein ratios
- 30
- 50
- 70
- 90
- 110
- 130
- 150
- 170
- 190
- 210
- For HRP make the following solutions to reflect an Au:Protein ratios
- 230
- 250
- 270
- 290
- 310
- 330
- 350
- 370
- 390
- 410
- Water up to 5mL
Figures
Notes
Stock Solutions
- Gold
- 49.3 mg HAuCl4·3H2O in 50mL water ---> 2.50mM
- HRP
- 6.6 mg HRP in 100 mL water ---> 1.5μM
- Myoglobin
- 28.7mg myoglobin in 100mL water ---> 16.9μM
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