User:Asya L. Tucker/Notebook/Asya 571/2015/09/02

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Objective

To make Lysozyme-AuNP fibers for future experiments.

Procedure

Dr. Hartings left a procedure for us to use in his lab notebook (also listed below).

Target Concentrations

Based on the total reaction volume that the class will be using and the final concentrations that will be in each reaction volume, I have determined that the following HAuCl4·H2O and Lysozyme masses are appropriate for today's stock solutions.

Stock solutions

  1. HAuCl4·H2O
    1. 2.5 mM
    2. MW = 339.79 g/mol
    3. Volume = 50 mL
    4. 42.4 mg
  2. Lysozyme
    1. 20 μM
    2. MW = 14307 g/mol
    3. Volume = 100 mL
    4. 28.6 mg

Actual Concentrations

The actual masses are different from the target masses. The concentrations shown below reflect the actual concentrations made for the stock solutions. I used a 50 mL volumetric flask to make the gold stock solution and a 100 mL volumetric flask to make the protein solution. In each case I used HPLC-grade water.

Stock solutions

  1. HAuCl4·H2O
    1. 50.32 mg
    2. MW = 339.79 g/mol
    3. Volume = 50 mL
    4. 2.96 mM
  2. Lysozyme
    1. 30.65 mg
    2. MW = 14307 g/mol
    3. Volume = 100 mL
    4. 21.4 μM

Directions for making samples

Note: The final concentration of gold should be 0.25 mM and the final concentration of lysozyme should be 5.556 μM. This gives a Au:lysozyme ratio of 45, which should yield fibers.

We prepared 16 # 1 mL samples

    1. Volume of gold stock: 84.4 μL
    2. Volume of protein stock: 259 μL
    3. Volume of water: 656 μL

We prepared 10 # 5 mL samples

    1. Volume of gold stock: 422 μL
    2. Volume of protein stock: 1,297 μL
    3. Volume of water: 3,281 μL

We placed the 16, 1 mL samples and the 10, 5 mL into the oven at 80 degrees Celsius for 4 hours.

Results

Dr. Hartings checked the samples after the oven cycle was complete and all of the reaction mixtures produced a homogeneous solution of nanoparticles (AuNPs) instead of fibers (AuNP fibers). The most likely reason is that the balance did not properly record the mass of the lysozyme. Dr. Harting prepared more; the procedure can be found here

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