User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2014/02/04

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Purpose/Objective

  • Synthesize lysozyme-AuNP.
  • Prepare stock solutions for all chelators.
  • Add the various chelators to lysozyme-AuNP solutions.
  • Change pH of lysozyme-AuNP using buffer solutions prepared yesterday.

Procedure

Chelators

Creating Stock Solutions of Chelators

1.29.data.chelator.png

  • 2,2 Bipyridine was used instead of 2, 2 bipyridyl. It stillhas the same function just a different chirality of the 2 nitrogen's.

Adding Chelators to Lysozyme-AuNP Solutions

  • Add 0.5 mL of each chelator stock solution to each 1.5 mL aliquot.
  • Record effects of each chelator on each solution.
  • Chelators are basic because NaOH had to be added to dissolve the respective chelators with the exception of 2,2 bipyridine.

1.29.data.chelator.add

  • 2,2 bipyridine was used instead of 2,2 bipyridyl

Note for all Chelators with the exception of 2,2 bipyridine, NaOH had to be added in order to get the chelator to dissolve in water. Each pH of the chelator were around a pH of 9-10. This was tested with pH strips.

pH

Adding Buffers to Lysozyme-AuNP Solutions

  1. Dilute the 1 M solutions to 100 mM, 10 mM, and 1 mM.
  2. Add 1 mL of each to the lysozyme-AuNP solutions.
  3. Let sit overnight and record results.

Made More AuNP

  • Because the 50:1 ratio did not form fibers as expected during our last round of synthesis we decided to make more AuNP's.
  • Specifically we made more 40:1 and 50:1 lysozyme lined AuNP's.
  • ASK CARLY FOR CALCULATIONS!!

Synthesizing Lysozyme-AuNP

Creating the Gold Stock Solution

  1. Add 0.0524 g of HAuCl4·3H2O to a 50 mL volumetric flask and fill with distilled water.
  2. The final concentration of gold stock was 2.66 mM.

Creating the Lysozyme Stock Solution

  1. Add 0.0717 g of lysozyme to a 50 mL volumetric flask and fill with distilled water.
  2. The final concentration of lysozyme stock was 100.28 μM.

Creating Au:Protein Ratios of 40:1 and 50:1

  1. For the 40:1 ratio, add 470 μL of gold solution, 312 μL of lysozyme solution, and 4218 μL of deionized water to a test tube.
  2. For the 50:1 ratio, add 470 μL of gold solution, 249 μL of lysozyme solution, and 4281 μL of deionized water to a test tube.
  3. We made 10 tubes of each ratio.
  4. The samples were heated at 80°C for 4 hours.