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

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(Protocol)
(HRP-AuNP and Pepsin-AuNP)
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| 210||0.496031746||1.19048E-06||1.19048E-06||0.389044507||0.379132545||4.114923747||4.124835709
 
| 210||0.496031746||1.19048E-06||1.19048E-06||0.389044507||0.379132545||4.114923747||4.124835709
 
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==Data==
 +
==UV/vis==
 +
*Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.
 +
<br>
 +
Figure 1. 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>
 +
Figure 2. Absorbance spectra of a 1/10 dilution of 2.52mM gold in water
 +
<br>
 +
[[Image:Golddilutionzem.png]]
 +
<br>
 +
Figure 3. Absorbance spectra of 1/2 dilution of 15.3μM HRP
 +
<br>
 +
[[Image:HalfdilutedHRP10222013zem.png]]
 +
<br>
 +
Figure 4. 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 14:23, 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

UV/vis

  • 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
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
Golddilutionzem.png
Figure 3. Absorbance spectra of 1/2 dilution of 15.3μM HRP
HalfdilutedHRP10222013zem.png
Figure 4. Absorbance spectrum of 1/10 dilution of 15.6μM pepsin
Pepsindilutionzem.png

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