- Measure pepsin activity of pepsin in solution and pepsin-AuNPs in solution
- Run UV-Vis on HRP and Pepsin linked Au-NP's synthesized yesterday
- Synthesize HRP- AuNP at higher ratios.
- Synthesize Myoglobin- AuNPs with the ratio used in yesterday's lab
- 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
- For HRP make the following solutions to reflect an Au:Protein ratios
- Water up to 5mL
- Place solutions in oven for 4 hours at 80 degrees Celsius
Calculations used to make solutions
- Purple tint of varying degrees:Lower ratios were magenta and higher ratios were blueish purple)
- No fibers formed
- 49.3 mg HAuCl4·3H2O in 50mL water ---> 2.50mM
- 6.6 mg HRP in 100 mL water ---> 1.5μM
- 28.7mg myoglobin in 100mL water ---> 16.9μM