User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/10/22

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==Notes==
==Notes==
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*The protocol for the synthesis of HRP-AuNP and Pepsin-AuNP can be found here [[User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/08/28]].
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<br>
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Table 1. Preparation of Protein-AuNP solutions
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''Amount of gold added(mL)'''
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| align="center" style="background:#f0f0f0;"|'''concentration HRP(M)'''
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| align="center" style="background:#f0f0f0;"|'''concentration Pepsin(M)'''
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| align="center" style="background:#f0f0f0;"|'''Volume HRP(mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Pepsin(mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Water HRP (mL)'''
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| align="center" style="background:#f0f0f0;"|'''Volume Water Pepsin (mL)'''
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|-
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| 30||0.496031746||8.33333E-06||8.33333E-06||2.723311547||2.653927813||1.780656707||1.850040441
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|-
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| 50||0.496031746||0.000005||0.000005||1.633986928||1.592356688||2.869981326||2.911611566
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|-
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| 70||0.496031746||3.57143E-06||3.57143E-06||1.16713352||1.137397634||3.336834734||3.36657062
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|-
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| 90||0.496031746||2.77778E-06||2.77778E-06||0.907770516||0.884642604||3.596197738||3.61932565
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|-
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| 110||0.496031746||2.27273E-06||2.27273E-06||0.742721331||0.723798494||3.761246923||3.780169759
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|-
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| 130||0.496031746||1.92308E-06||1.92308E-06||0.628456511||0.61244488||3.875511743||3.891523374
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|-
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| 150||0.496031746||1.66667E-06||1.66667E-06||0.544662309||0.530785563||3.959305945||3.973182691
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|-
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| 170||0.496031746||1.47059E-06||1.47059E-06||0.480584391||0.468340202||4.023383863||4.035628052
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|-
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| 190||0.496031746||1.31579E-06||1.31579E-06||0.42999656||0.419041234||4.073971694||4.08492702
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|-
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| 210||0.496031746||1.19048E-06||1.19048E-06||0.389044507||0.379132545||4.114923747||4.124835709
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|}
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<br>
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*The varying ratios will be used to determine the ideal concentrations for the preparation of fibers of unraveled proteins.
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<br>
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*Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.
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<br>
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Figure 1. Absorbance spectra of a 1/10 dilution of BSA-AuNPs
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<br>
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[[Image:BSAAuNPsdilutionzem.png]]
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<br>
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*Absorbance of all starting materials used for synthesis of AuNPs today for future comparison
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<br>
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Figure 2. Absorbance spectra of a 1/10 dilution of 2.52mM gold in water
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<br>
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[[Image:Golddilutionzem.png]]
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<br>
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Figure 3. Absorbance spectra of 1/2 dilution of 15.3μM HRP
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<br>
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[[Image:HalfdilutedHRP10222013zem.png]]
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<br>
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Figure 4. Absorbance spectrum of 1/10 dilution of 15.6μM pepsin
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<br>
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[[Image:Pepsindilutionzem.png]]
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<br>
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*No absorbance peak was observed for pepsin.

Current revision

Project name Main project page
Previous entry      Next entry

Objective

  1. Synthesize Pepsin-AuNPs for reactivity tests tomorrow
  2. Determine the Au:Protein ratio required for fiber formation for pepsin and HRP

Description

  1. Synthesis of HRP-AuNPs and Pepsin-AuNPs
    1. Into test tube that will contain a total of 5mL add
    2. Au solution such that the final concentration is 0.25mM
    3. Protein solution such that the final concentration reflects the following Au:Protein ratios
      1. 30
      2. 50
      3. 70
      4. 90
      5. 110
      6. 130
      7. 150
      8. 170
      9. 190
      10. 210
    4. Water up to 5mL

Data

Stock Solutions

  1. HRP
    1. 67.3mg in 100mL of water ---> 15.3 μM
  2. Pepsin
    1. 54.4mg in 100mL of water ---> 15.7 μM
  3. Gold
    1. Need 50mL of 2.5mM
    2. 49.7mg of HAuCl4·3H2O in 50mL water --> 2.52 mM

Notes


Table 1. Preparation of Protein-AuNP solutions

Ratio Amount of gold added(mL) concentration HRP(M) concentration Pepsin(M) Volume HRP(mL) Volume Pepsin(mL) Volume Water HRP (mL) Volume Water Pepsin (mL)
300.4960317468.33333E-068.33333E-062.7233115472.6539278131.7806567071.850040441
500.4960317460.0000050.0000051.6339869281.5923566882.8699813262.911611566
700.4960317463.57143E-063.57143E-061.167133521.1373976343.3368347343.36657062
900.4960317462.77778E-062.77778E-060.9077705160.8846426043.5961977383.61932565
1100.4960317462.27273E-062.27273E-060.7427213310.7237984943.7612469233.780169759
1300.4960317461.92308E-061.92308E-060.6284565110.612444883.8755117433.891523374
1500.4960317461.66667E-061.66667E-060.5446623090.5307855633.9593059453.973182691
1700.4960317461.47059E-061.47059E-060.4805843910.4683402024.0233838634.035628052
1900.4960317461.31579E-061.31579E-060.429996560.4190412344.0739716944.08492702
2100.4960317461.19048E-061.19048E-060.3890445070.3791325454.1149237474.124835709


  • The varying ratios will be used to determine the ideal concentrations for the preparation of fibers of unraveled proteins.


  • Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.


Figure 1. Absorbance spectra of a 1/10 dilution of BSA-AuNPs
Image:BSAAuNPsdilutionzem.png

  • Absorbance of all starting materials used for synthesis of AuNPs today for future comparison


Figure 2. Absorbance spectra of a 1/10 dilution of 2.52mM gold in water
Image:Golddilutionzem.png
Figure 3. Absorbance spectra of 1/2 dilution of 15.3μM HRP
Image:HalfdilutedHRP10222013zem.png
Figure 4. Absorbance spectrum of 1/10 dilution of 15.6μM pepsin
Image:Pepsindilutionzem.png

  • No absorbance peak was observed for pepsin.



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