Objectives
- Run UV-vis on the 20 protein-Au nanoparticle samples prepared on 10/22/2013.
- Synthesize new HRP-AuNP solutions and Myoglobin-AuNP solutions for future catalytic activity testing.
HRP-AuNP and Myoglobin-AuNP
- The protein-AuNP solutions prepared on 10/22/2013 did not produce fibers. As such, new protein-AuNP solutions with increased ratios of protein-AuNP were prepared in order to determine the optimal concentration of Au to protein to synthesize fibers.
- The general protocol for the synthesis of HRP-AuNP and Pepsin-AuNP follows the general protocol for the preparation of protein wrapped Au-NPs described on 08/28/2013.
- 5mL of each solution was prepared.
Table 1. Stock solutions used in the preparation of Protein-AuNP solutions
Concentration HRP Day 2(M)
|
Concentration Gold Stock (M)
|
Myoglobin (M)
|
0.0000015 |
0.0025 |
0.0000165
|
Table 2. Preparation of HRP-AuNP solutions
Ratio
|
Amount of gold added (mL)
|
Concentration HRP (M)
|
Volume HRP (mL)
|
Volume H2O added (mL)
|
230 |
0.5 |
1.08696E-06 |
3.623188406 |
0.876811594
|
250 |
0.5 |
0.000001 |
3.333333333 |
1.166666667
|
270 |
0.5 |
9.25926E-07 |
3.086419753 |
1.413580247
|
290 |
0.5 |
8.62069E-07 |
2.873563218 |
1.626436782
|
310 |
0.5 |
8.06452E-07 |
2.688172043 |
1.811827957
|
330 |
0.5 |
7.57576E-07 |
2.525252525 |
1.974747475
|
350 |
0.5 |
7.14286E-07 |
2.380952381 |
2.119047619
|
370 |
0.5 |
6.75676E-07 |
2.252252252 |
2.247747748
|
390 |
0.5 |
6.41026E-07 |
2.136752137 |
2.363247863
|
410 |
0.5 |
6.09756E-07 |
2.032520325 |
2.467479675
|
Table 3. Preparation of Myoglobin-AuNPs
Ratio
|
Amount of gold added (mL)
|
Concentration Myoglobin (M)
|
Volume of myogloblin(mL)
|
Volume Water Myoglobin(mL)
|
30 |
0.5 |
8.33333E-06 |
2.525252525 |
1.974747475
|
50 |
0.5 |
0.000005 |
1.515151515 |
2.984848485
|
70 |
0.5 |
3.57143E-06 |
1.082251082 |
3.417748918
|
90 |
0.5 |
2.77778E-06 |
0.841750842 |
3.658249158
|
110 |
0.5 |
2.27273E-06 |
0.688705234 |
3.811294766
|
130 |
0.5 |
1.92308E-06 |
0.582750583 |
3.917249417
|
150 |
0.5 |
1.66667E-06 |
0.505050505 |
3.994949495
|
170 |
0.5 |
1.47059E-06 |
0.445632799 |
4.054367201
|
190 |
0.5 |
1.31579E-06 |
0.398724083 |
4.101275917
|
210 |
1.19048E-06 |
0.360750361 |
4.139249639
|
UV/vis of Pepsin-AuNP and HRP-AuNP
General Protocol:
- Place 1mL of each ratio in an 1.5mL eppendorf tube.
- Centrifuge at maximum speed for 30 minutes.
- Take a UV-Vis spectrum of each sample from 200nm to 800nm.
- After taking the spectrum, store the sample in a fresh eppy tube in an upright position.
Figure 1. Corrected Absorbance spectra of pepsin-AuNPs
- Both 30:1 and 70:1 pepsin-AuNP had an absorbance peak at 518nm.
Notes
- The remaining UV/vis samples will be run during the next lab period.
- Although neither the HRP or the pepsin solutions formed fibers, a black residue and color change(see figure 2) was observed for the pepsin solutions. The black residue will be tested in the future for identification.
|