Difference between revisions of "User:Moira M. Esson/Notebook/CHEM-571/2013/10/23"

From OpenWetWare
Jump to: navigation, search
(HRP-AuNP and Myoglobin-AuNP)
(UV/vis of Pepsin-AuNP and HRP-AuNP)
Line 86: Line 86:
 
<br>
 
<br>
 
==UV/vis of Pepsin-AuNP and HRP-AuNP==
 
==UV/vis of Pepsin-AuNP and HRP-AuNP==
The general protocol for the UV/vis spectrum on the prepared protein-AuNP solutions was follwed
+
'''General Protocol''':
 
# Place 1mL of each ratio in an 1.5mL eppendorf tube.  
 
# Place 1mL of each ratio in an 1.5mL eppendorf tube.  
 
# Centrifuge at maximum speed for 30 minutes.
 
# Centrifuge at maximum speed for 30 minutes.
 
# Take a UV-Vis spectrum of each sample from 200nm to 800nm.
 
# 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.
 
# 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>
 +
[[
 +
<br>
 +
*Both 30:1 and 70:1 pepsin-AuNP had an absorbance peak at 518nm.
 +
 
==Notes==
 
==Notes==
 
*The remaining UV/vis samples will be run during the next lab period.
 
*The remaining UV/vis samples will be run during the next lab period.

Revision as of 18:41, 27 October 2013

Owwnotebook icon.png Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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
[[

  • 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