User:Moira M. Esson/Notebook/CHEM-571/2013/11/06

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(UV/vis Analysis)
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==Entry title==
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==Objectives==
#Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
#Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
#Analyze protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using UV/vis spectroscopy.
#Analyze protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using UV/vis spectroscopy.
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==UV/vis Analysis==
==UV/vis Analysis==
'''General Protocol''':
'''General Protocol''':

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Objectives

  1. Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
  2. Analyze protein-AuNP solutions prepared on 10/30/2013 using UV/vis spectroscopy.


UV/vis Analysis

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 Lysozyme-AuNP solutions

Image:Corrected Absorbance Spectra of Lysozyme-AuNP solutions.png

  • Only the 30:1 ratio exhibited an absorbance peak at 526nm. This was expected as this was the only colored solution or only solution that did not form fibers shown on 11/05/2013


Figure 2. Corrected Absorbance Spectra of Myoglobin Au-NP solutions
Image:Corrected Absorbance spectra of Myoglobin Au-NP solution .png

  • 15:1 ratio is the only exhibited an aborbance peak at 732nm. This may indicate the optimal concentration for the preparation of fibers. Although this solution produced fibers 11/05/2013 this solution is still easily monitored using UV/vis spectroscopy.

Protein-AuNP solution preparation

  • The general protocol for the synthesis of Hemoglobin-AuNP and Lysozyme-AuNP follows the general protocol for the preparation of protein wrapped Au-NPs described on 08/28/2013.
  • 5mL of each solution was prepared.
  • All solutions were prepared using deionized H2O
  • Tri-hydrate gold was used for the preparation of the gold standard solution.
  • All stock solutions were prepared in a 10mL volumetric flask.


Table 1. Preparation of Hemoglobin Stock solution

Molecular weight (g/mol) 64500
mass myoglobin (g)0.0106
volume solution (L)0.01
Concentration (M)1.64341E-05


Table 2. Preparation of Gold stock solution

Molecular weight Au (g/mol) 393.83
mass Au(g)0.032
volume stock(L)0.025
Concentration (M)0.003250133


Table 3. Preparation of Lysozyme stock solution

Molecular weight (g/mol) 14307
mass lysozyme (g)0.0717
volume solution (L)0.05
Concentration (M)0.00010028


Table 4. Preparation of Hemoglobin-AuNP solutions

Ratio Amount of gold added concentration hemoglobin (M) volume Hemoglobin (mL) Volume Water (mL)
300.3845996098.33333E-062.5353773582.080023032
500.3845996090.0000051.5212264153.094173976
700.3845996093.57143E-061.0865902963.528810094
900.3845996092.77778E-060.8451257863.770274604
1100.3845996092.27273E-060.6914665523.923933838
1300.3845996091.92308E-060.5850870834.030313308
1500.3845996091.66667E-060.5070754724.108324919
1700.3845996091.47059E-060.4474195344.167980857
1900.3845996091.31579E-060.4003227414.21507765
2100.3845996091.19048E-060.3621967654.253203625


Table 5. Preparation of Lysozyme Au-NP solutions

Ratio Amount gold added (mL) Concentration Lysozyme (M) Volume lysozyme (mL) Volume water (mL)
50.3845996090.000052.4930195452.122380845
100.3845996090.0000251.2465097733.368890618
150.3845996091.66667E-050.8310065153.784393876
200.3845996090.00001250.6232548863.992145504
250.3845996090.000010.4986039094.116796482
300.3845996098.33333E-060.4155032584.199897133
350.3845996097.14286E-060.3561456494.259254741
400.3845996090.000006250.3116274434.303772947
450.3845996095.55556E-060.2770021724.338398219
500.3845996090.0000050.2493019554.366098436
  • New ratios of lysozyme were prepared in order to determine the optimal concentration ratios for the formation of fibers.


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