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

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Current revision (17:24, 29 October 2013) (view source)
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*Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.
*Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.
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Absorbance spectra of a 1/10 dilution of BSA-AuNPs
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Absorbance spectra of a 1/10 dilution of BSA-AuNPs
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[[Image:BSAAuNPsdilutionzem.png]]
[[Image:BSAAuNPsdilutionzem.png]]

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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)
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


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
Image:BSAAuNPsdilutionzem.png

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

  • 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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