User:Moira M. Esson/Notebook/CHEM-571/2013/10/23

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(UV/vis of Pepsin-AuNP and HRP-AuNP)
(Notes)
Line 100: Line 100:
==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.
-
*Although  
+
*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.
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Revision as of 22:48, 27 October 2013

Project name Main project page
Previous entry      Next entry

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)
0.00000150.00250.0000165


Table 2. Preparation of HRP-AuNP solutions

Ratio Amount of gold added (mL) Concentration HRP (M) Volume HRP (mL) Volume H2O added (mL)
2300.51.08696E-063.6231884060.876811594
2500.50.0000013.3333333331.166666667
2700.59.25926E-073.0864197531.413580247
2900.58.62069E-072.8735632181.626436782
3100.58.06452E-072.6881720431.811827957
3300.57.57576E-072.5252525251.974747475
3500.57.14286E-072.3809523812.119047619
3700.56.75676E-072.2522522522.247747748
3900.56.41026E-072.1367521372.363247863
4100.56.09756E-072.0325203252.467479675


Table 3. Preparation of Myoglobin-AuNPs

Ratio Amount of gold added (mL) Concentration Myoglobin (M) Volume of myogloblin(mL) Volume Water Myoglobin(mL)
300.58.33333E-062.5252525251.974747475
500.50.0000051.5151515152.984848485
700.53.57143E-061.0822510823.417748918
900.52.77778E-060.8417508423.658249158
1100.52.27273E-060.6887052343.811294766
1300.51.92308E-060.5827505833.917249417
1500.51.66667E-060.5050505053.994949495
1700.51.47059E-060.4456327994.054367201
1900.51.31579E-060.3987240834.101275917
2101.19048E-060.3607503614.139249639


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 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.


Personal tools