User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/10/23

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Current revision (00:06, 30 November 2012) (view source)
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''Preparing Solutions for Chemilluminescence''
''Preparing Solutions for Chemilluminescence''
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*1mM sodium carbonate solution and 1mM sodium bicarbonate solution were prepared in water. The pH of the 1mM sodium carbonate solution was adjusted to 8.5 by adding 1mM sodium bicarbonate solution, drop-wise. The final concentration of the sodium carbonate buffer was calculated to be 0.833mM.
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*0.564 mM luminol stock solution was prepared in carbonate buffer
*0.564 mM luminol stock solution was prepared in carbonate buffer
**0.0010 g luminol + 10 mL carbonate buffer --> 0.564 mM luminol solution.
**0.0010 g luminol + 10 mL carbonate buffer --> 0.564 mM luminol solution.

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Objective

  • to create protein fibers in lysozyme/Au solutions
  • to prepare solutions for chemiluminescence reactions

Description

Lysozyme/Au Solutions

  • HAuCl4 and Lysozyme solutions were prepared.
    • 7.4mg HAuCl4 in 5.0 mL --> 4.36 mM
    • 0.001g Lysozyme in 5.0 mL -->5mM
    • 8mL mixtures with mole ratios of 10, 15, 20, 25, 30, 35, and 40 Au/Lysozyme were made and microwaved for 2 minutes, based on a protocol from the paper Cytotoxicity and cellular uptake of lysozyme-stabilized gold nanoparticles by Yeonju Lee and Kurt E. Geckeler.
      • The solutions bubbled over in the microwave. No fibers were formed. Next time, will try the same protocol as used when creating the Au/BSA mixtures in September.


Preparing Solutions for Chemilluminescence

  • 1mM sodium carbonate solution and 1mM sodium bicarbonate solution were prepared in water. The pH of the 1mM sodium carbonate solution was adjusted to 8.5 by adding 1mM sodium bicarbonate solution, drop-wise. The final concentration of the sodium carbonate buffer was calculated to be 0.833mM.
  • 0.564 mM luminol stock solution was prepared in carbonate buffer
    • 0.0010 g luminol + 10 mL carbonate buffer --> 0.564 mM luminol solution.
    • Goal: 0.5mM solution
  • 5 mM H2O2 stock solution in carbonate buffer:
    • 3.9 μL of 30 wt% H2O2 + 10 mL of carbonate buffer --> 5 mM H2O2solution.
  • 9.2 μM HRP solution
    • 4.0 mg HRP + 1 mL autoclaved water --> 9.2 μM HRP solution.


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