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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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*The protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/22|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 protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/22|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 [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]].
 
*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 [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]].
 +
*5mL of each solution was prepared.
 
<br>
 
<br>
 
Table 1. Stock solutions used in the preparation of Protein-AuNP solutions
 
Table 1. Stock solutions used in the preparation of Protein-AuNP solutions
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| 0.0000015||0.0025||0.0000165
 
| 0.0000015||0.0025||0.0000165
 
|}  
 
|}  
Table 2. Preparation of Protein-AuNP solutions
+
<br>
 +
Table 2. Preparation of HRP-AuNP solutions
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 +
| align="center" style="background:#f0f0f0;"|'''Amount of gold added (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''Concentration HRP (M)'''
 +
| align="center" style="background:#f0f0f0;"|'''Volume HRP (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''Volume H<sub>2</sub>O 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
 +
|}
 +
<br>
 +
Table 3. Preparation of Myoglobin-AuNPs
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 +
| align="center" style="background:#f0f0f0;"|'''Amount of gold added (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''Concentration Myoglobin (M)'''
 +
| align="center" style="background:#f0f0f0;"|'''Volume of myogloblin(mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''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
 +
|}
 +
<br>
 +
 
 +
==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.
 +
<br>
 +
Figure 1. Corrected Absorbance spectra of pepsin-AuNPs
 +
<br>
 +
[[Image:Pepsin Nano.png]]
 +
<br>
 +
*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.
  
 
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Latest revision as of 22:30, 26 September 2017

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Objectives

  1. Run UV-vis on the 20 protein-Au nanoparticle samples prepared on 10/22/2013.
  2. 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:

  1. Place 1mL of each ratio in an 1.5mL eppendorf tube.
  2. Centrifuge at maximum speed for 30 minutes.
  3. Take a UV-Vis spectrum of each sample from 200nm to 800nm.
  4. After taking the spectrum, store the sample in a fresh eppy tube in an upright position.


Figure 1. Corrected Absorbance spectra of pepsin-AuNPs
Pepsin Nano.png

  • 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.