User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05

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(Procedure)
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*# The tris buffer and Au/BSA solution were mixed gently and let sit for 30 minutes.  
*# The tris buffer and Au/BSA solution were mixed gently and let sit for 30 minutes.  
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*# s
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*# After 30 minutes, 500uL of each sample were loaded into quark cuvette and placed into UV-Vis to measure absorbance from 800nm to 200mm.
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*# Graph were plotted with Absorbance versus Wavelength, for details of absorbance from each sample, see [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/05|Dhea Patel's Lab Notebook]]
==Notes==
==Notes==

Revision as of 15:42, 8 September 2012

Experimental Biological Chemistry I Main project page
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Purpose

  • To resuspend gold nano-particle pallets with Tris buffer.
  • To determine the best concentration and pH of Tris buffer for resuspension via measuring the absorbance of resuspended solutions throughout time.
  • To make new sets of Au/BSA solutions with varies ratios.

Procedure

  • A new sets of solutions of BSA and HAuCl4 mixture were made. New stock solutions of HAuCl4 and BSA were also made.
  • 100mM tris buffer at pH 8.0 and pH 10.0 were diluted with serial dilution to four different concentrations: 10mM, 1mM, 100uM, and 10uM, respectively.
  • Resuspending Au/BSA solutions in Tris buffer
    1. The old sets of solutions of Au/BSA at different ratios were spin in centrifuge at 3000rpm for 5 minutes at room temperature.
    2. The supernatant were poured out. Pallet of different Au/BSA ratios were resuspended in different concentrations and different pH of Tris buffer.
    3. The details of buffer-pallet mix is shown in table below:
Ratio of Au/BSA Pallet Tris Buffer Concentrations [mM] Tris Buffer pH
132 10 10
133 1 10
134 0.1 10
136 0.01 10
138 10 8
140 1 8
160 0.1 8
170 0.01 8
    1. The tris buffer and Au/BSA solution were mixed gently and let sit for 30 minutes.
    2. After 30 minutes, 500uL of each sample were loaded into quark cuvette and placed into UV-Vis to measure absorbance from 800nm to 200mm.
    3. Graph were plotted with Absorbance versus Wavelength, for details of absorbance from each sample, see Dhea Patel's Lab Notebook

Notes

  • The new sets of solutions made on 2012/09/04 failed to react. The solutions after heating appeared yellow in color. This indicates the BSA failed to react with the HAuCl4 in solution.



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