User:Eleni N. Kalivas/Notebook/CHEM-571/2013/10/22: Difference between revisions

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### 210
### 210
## Water up to 5mL
## Water up to 5mL
==HRP-AuNP and Pepsin-AuNP==
<br>
Table 1. Preparation of Protein-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;"|'''concentration Pepsin(M)'''
| align="center" style="background:#f0f0f0;"|'''Volume HRP(mL)'''
| align="center" style="background:#f0f0f0;"|'''Volume Pepsin(mL)'''
| align="center" style="background:#f0f0f0;"|'''Volume Water HRP (mL)'''
| align="center" style="background:#f0f0f0;"|'''Volume Water Pepsin (mL)'''
|-
| 30||0.496031746||8.33333E-06||8.33333E-06||2.723311547||2.653927813||1.780656707||1.850040441
|-
| 50||0.496031746||0.000005||0.000005||1.633986928||1.592356688||2.869981326||2.911611566
|-
| 70||0.496031746||3.57143E-06||3.57143E-06||1.16713352||1.137397634||3.336834734||3.36657062
|-
| 90||0.496031746||2.77778E-06||2.77778E-06||0.907770516||0.884642604||3.596197738||3.61932565
|-
| 110||0.496031746||2.27273E-06||2.27273E-06||0.742721331||0.723798494||3.761246923||3.780169759
|-
| 130||0.496031746||1.92308E-06||1.92308E-06||0.628456511||0.61244488||3.875511743||3.891523374
|-
| 150||0.496031746||1.66667E-06||1.66667E-06||0.544662309||0.530785563||3.959305945||3.973182691
|-
| 170||0.496031746||1.47059E-06||1.47059E-06||0.480584391||0.468340202||4.023383863||4.035628052
|-
| 190||0.496031746||1.31579E-06||1.31579E-06||0.42999656||0.419041234||4.073971694||4.08492702
|-
| 210||0.496031746||1.19048E-06||1.19048E-06||0.389044507||0.379132545||4.114923747||4.124835709
|}
==Data==
*Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.
<br>
Absorbance spectra of a 1/10 dilution of BSA-AuNPs
<br>
[[Image:BSAAuNPsdilutionzem.png]]
<br>
*Absorbance of all starting materials used for synthesis of AuNPs today for future comparison
<br>
Absorbance spectra of a 1/10 dilution of 2.52mM gold in water
<br>
[[Image:Golddilutionzem.png]]
<br>
Absorbance spectra of 1/2 dilution of 15.3μM HRP
<br>
[[Image:HalfdilutedHRP10222013zem.png]]
<br>
Absorbance spectrum of 1/10 dilution of 15.6μM pepsin
<br>
[[Image:Pepsindilutionzem.png]]
<br>
*No absorbance peak was observed for pepsin. This was expected due to the lack of color of the pepsin stock solution.




Revision as of 14:24, 29 October 2013

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Objective

The objective of today's lab is to synthesize AuNPs for reactivity tests tomorrow. Also to determine the Au;Protein ratio that is required for fiber formation of HRP and for pepsin.

Protocol

The following has been taken from Matt Harting's lab notebook.

  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


HRP-AuNP and Pepsin-AuNP


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)
30 0.496031746 8.33333E-06 8.33333E-06 2.723311547 2.653927813 1.780656707 1.850040441
50 0.496031746 0.000005 0.000005 1.633986928 1.592356688 2.869981326 2.911611566
70 0.496031746 3.57143E-06 3.57143E-06 1.16713352 1.137397634 3.336834734 3.36657062
90 0.496031746 2.77778E-06 2.77778E-06 0.907770516 0.884642604 3.596197738 3.61932565
110 0.496031746 2.27273E-06 2.27273E-06 0.742721331 0.723798494 3.761246923 3.780169759
130 0.496031746 1.92308E-06 1.92308E-06 0.628456511 0.61244488 3.875511743 3.891523374
150 0.496031746 1.66667E-06 1.66667E-06 0.544662309 0.530785563 3.959305945 3.973182691
170 0.496031746 1.47059E-06 1.47059E-06 0.480584391 0.468340202 4.023383863 4.035628052
190 0.496031746 1.31579E-06 1.31579E-06 0.42999656 0.419041234 4.073971694 4.08492702
210 0.496031746 1.19048E-06 1.19048E-06 0.389044507 0.379132545 4.114923747 4.124835709


Data

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


Absorbance spectra of a 1/10 dilution of BSA-AuNPs

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


Absorbance spectra of a 1/10 dilution of 2.52mM gold in water

Absorbance spectra of 1/2 dilution of 15.3μM HRP

Absorbance spectrum of 1/10 dilution of 15.6μM pepsin

  • No absorbance peak was observed for pepsin. This was expected due to the lack of color of the pepsin stock solution.



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