Difference between revisions of "User:Trisha I. Ibeh/Notebook/Trisha Notebook/2013/10/23"

Biomaterials Design Lab <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Objective

• 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

Procedure

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
6. Place solutions in oven for 4 hours at 80 degrees Celsius

Calculations used to make solutions

Notes

Observations

• Purple tint of varying degrees:Lower ratios were magenta and higher ratios were blueish purple)
• No fibers formed
• Fibers not formed

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